Compositions useful as inhibitors of protein kinases

ABSTRACT

The present invention relates to compounds useful as inhibitors of protein kinases. The invention also provides processes for preparing the compounds, pharmaceutically acceptable compositions comprising the compounds, and methods of using the compounds and compositions in the treatment of various disease, conditions, or disorders.

CROSS-REFERENCE TO RELATED APPLICATION(S)

This application is a continuation of U.S. patent application Ser. No.10/965,695, filed Oct. 14, 2004 (issued as U.S. Pat. No. 7,700,609),which application claims priority to U.S. Provisional Patent ApplicationSer. No. 60/510,881, filed Oct. 14, 2003. The entire disclosures ofthese prior applications are incorporated herein by reference.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to compounds useful as inhibitors ofprotein kinases. The invention also provides processes for preparing thecompounds of the invention, pharmaceutically acceptable compositionscomprising the compounds, and methods of using the compounds andcompositions in the treatment of various disorders.

BACKGROUND OF THE INVENTION

The search for new therapeutic agents has been greatly aided in recentyears by a better understanding of the structure of enzymes and otherbiomolecules associated with diseases. One important class of enzymesthat has been the subject of extensive study is protein kinases.

Protein kinases constitute a large family of structurally relatedenzymes that are responsible for the control of a variety of signaltransduction processes within the cell. (See, Hardie, G. and Hanks, S.The Protein Kinase Facts Book, I and II, Academic Press, San Diego,Calif.: 1995). Protein kinases are thought to have evolved from a commonancestral gene due to the conservation of their structure and catalyticfunction. Almost all kinases contain a similar 250-300 amino acidcatalytic domain. The kinases may be categorized into families by thesubstrates they phosphorylate (e.g., protein-tyrosine,protein-serine/threonine, lipids, etc.). Sequence motifs have beenidentified that generally correspond to each of these kinase families(See, for example, Hanks, S. K., Hunter, T., FASEB J. 1995, 9, 576-596;Knighton et al., Science 1991, 253, 407-414; Hiles et al., Cell 1992,70, 419-429; Kunz et al., Cell 1993, 73, 585-596; Garcia-Bustos et al.,EMBO J. 1994, 13, 2352-2361).

In general, protein kinases mediate intracellular signaling by effectinga phosphoryl transfer from a nucleoside triphosphate to a proteinacceptor that is involved in a signaling pathway. These phosphorylationevents act as molecular on/off switches that can modulate or regulatethe target protein biological function. These phosphorylation events areultimately triggered in response to a variety of extracellular and otherstimuli. Examples of such stimuli include environmental and chemicalstress signals (e.g., osmotic shock, heat shock, ultraviolet radiation,bacterial endotoxin, and H₂O₂), cytokines (e.g., interleukin-1 (IL-1)and tumor necrosis factor α (TNF-α)), and growth factors (e.g.,granulocyte macrophage-colony-stimulating factor (GM-CSF), andfibroblast growth factor (FGF)). An extracellular stimulus may affectone or more cellular responses related to cell growth, migration,differentiation, secretion of hormones, activation of transcriptionfactors, muscle contraction, glucose metabolism, control of proteinsynthesis, and regulation of the cell cycle.

Many diseases are associated with abnormal cellular responses triggeredby protein kinase-mediated events as described above. These diseasesinclude, but are not limited to, autoimmune diseases, inflammatorydiseases, bone diseases, metabolic diseases, neurological andneurodegenerative diseases, cancer, cardiovascular diseases, allergiesand asthma, Alzheimer's disease, and hormone-related diseases.Accordingly, there has been a substantial effort in medicinal chemistryto find protein kinase inhibitors that are effective as therapeuticagents.

The Aurora family of serine/threonine kinases is essential for cellproliferation [Bischoff, J. R. & Plowman, G. D. (The Aurora/Ipl1p kinasefamily: regulators of chromosome segregation and cytokinesis) Trends inCell Biology 9, 454-459 (1999); Giet, R. and Prigent, C.(Aurora/Ipl1p-related kinases, a new oncogenic family of mitoticserine-threonine kinases) Journal of Cell Science 112, 3591-3601 (1999);Nigg, E. A. (Mitotic kinases as regulators of cell division and itscheckpoints) Nat. Rev. Mol. Cell Biol. 2, 21-32 (2001); Adams, R. R,Carmena, M., and Earnshaw, W. C. (Chromosomal passengers and the(aurora) ABCs of mitosis) Trends in Cell Biology 11, 49-54 (2001)].Inhibitors of the Aurora kinase family therefore have the potential toblock growth of all tumour types.

The three known mammalian family members, Aurora-A (“2”), B (“1”) and C(“3”), are highly homologous proteins responsible for chromosomesegregation, mitotic spindle function and cytokinesis. Aurora expressionis low or undetectable in resting cells, with expression and activitypeaking during the G2 and mitotic phases in cycling cells. In mammaliancells proposed substrates for Aurora include histone H3, a proteininvolved in chromosome condensation, and CENP-A, myosin II regulatorylight chain, protein phosphatase 1, TPX2, all of which are requiredforcell division.

Since its discovery in 1997 the mammalian Aurora kinase family has beenclosely linked to tumorigenesis. The most compelling evidence for thisis that over-expression of Aurora-A transforms rodent fibroblasts(Bischoff, J. R., et al. A homologue of Drosophila aurora kinase isoncogenic and amplified in human colorectal cancers. EMBO J. 17,3052-3065 (1998)). Cells with elevated levels of this kinase containmultiple centrosomes and multipolar spindles, and rapidly becomeaneuploid. The oncogenic activity of Aurora kinases is likely to belinked to the generation of such genetic instability. Indeed, acorrelation between amplification of the aurora-A locus and chromosomalinstability in mammary and gastric tumours has been observed. (Miyoshi,Y., Iwao, K., Egawa, C., and Noguchi, S. Association of centrosomalkinase STK15/BTAK mRNA expression with chromosomal instability in humanbreast cancers. Int. J. Cancer 92, 370-373 (2001). (Sakakura, C. et al.Tumor-amplified kinase BTAK is amplified and overexpressed in gastriccancers with possible involvement in aneuploid formation. BritishJournal of Cancer 84, 824-831 (2001)). The Aurora kinases have beenreported to be over-expressed in a wide range of human tumours. Elevatedexpression of Aurora-A has been detected in over 50% of colorectalcancers (Bischoff, J. R., et al. A homologue of Drosophila aurora kinaseis oncogenic and amplified in human colorectal cancers. EMBO J. 17,3052-3065 (1998)) (Takahashi, T., et al. Centrosomal kinases, HsAIRk1and HsAIRK3, are overexpressed in primary colorectal cancers. Jpn. J.Cancer Res. 91, 1007-1014 (2000)), ovarian cancers (Gritsko, T. M. etal. Activation and overexpression of centrosome kinase BTAK/Aurora-A inhuman ovarian cancer. Clinical Cancer Research 9, 1420-1426 (2003)), andgastric tumors (Sakakura, C. et al. Tumor-amplified kinase BTAK isamplified and overexpressed in gastric cancers with possible involvementin aneuploid formation. British Journal of Cancer 84, 824-831 (2001)),and in 94% of invasive duct adenocarcinomas of the breast (Tanaka, T.,et al. Centrosomal kinase AIK1 is overexpressed in invasive ductalcarcinoma of the breast. Cancer Research. 59, 2041-2044 (1999)). Highlevels of Aurora-A have also been reported in renal, cervical,neuroblastoma, melanoma, lymphoma, pancreatic and prostate tumour celllines. (Bischoff, J. R., et al. A homologue of Drosophila aurora kinaseis oncogenic and amplified in human colorectal cancers. EMBO J. 17,3052-3065 (1998) (Kimura, M., Matsuda, Y., Yoshioka, T., and Okano, Y.Cell cycle-dependent expression and centrosomal localization of a thirdhuman Aurora/Ipl1-related protein kinase, AIK3. Journal of BiologicalChemistry 274, 7334-7340 (1999)) (Zhou et al. Tumour amplified kinaseSTK15/BTAK induces centrosome amplification, aneuploidy andtransformation Nature Genetics 20: 189-193 (1998)) (Li et al.Overexpression of oncogenic STK15/BTAK/Aurora-A kinase in humanpancreatic cancer Clin Cancer Res. 9(3):991-7 (2003)).Amplification/overexpression of Aurora-A is observed in human bladdercancers and amplification of Aurora-A is associated with aneuploidy andaggressive clinical behaviour (Sen S. et al.“Amplification/overexpression of a mitotic kinase gene in human bladdercancer” J. Natl. Cancer Inst. 94(17):1320-9 (2002)). Moreover,amplification of the aurora-A locus (20q13) correlates with poorprognosis for patients with node-negative breast cancer (Isola, J. J.,et al. “Genetic aberrations detected by comparative genomichybridization predict outcome in node-negative breast cancer” AmericanJournal of Pathology 147, 905-911 (1995)). Aurora-B is highly expressedin multiple human tumour cell lines, including leukemic cells (Katayamaet al. Human AIM-1: cDNA cloning and reduced expression duringendomitosis in megakaryocyte-lineage cells. Gene 244:1-7)). Levels ofthis enzyme increase as a function of Duke's stage in primary colorectalcancers (Katayama, H. et al. Mitotic kinase expression and colorectalcancer progression. Journal of the National Cancer Institute 91,1160-1162 (1999)). Aurora-C, which is normally only found in germ cells,is also over-expressed in a high percentage of primary colorectalcancers and in a variety of tumour cell lines including cervicaladenocarinoma and breast carcinoma cells (Kimura, M., Matsuda, Y.,Yoshioka, T., and Okano, Y. Cell cycle-dependent expression andcentrosomal localization of a third human Aurora/Ipl1-related proteinkinase, AIK3. Journal of Biological Chemistry 274, 7334-7340 (1999).(Takahashi, T., et al. Centrosomal kinases, HsAIRk1 and HsAIRK3, areoverexpressed in primary colorectal cancers. Jpn. J. Cancer Res. 91,1007-1014 (2000)).

Based on the known function of the Aurora kinases, inhibition of theiractivity should disrupt mitosis leading to cell cycle arrest. In vivo,an Aurora inhibitor therefore slows tumor growth and induces regression.

Elevated levels of all Aurora family members are observed in a widevariety of tumour cell lines. Aurora kinases are over-expressed in manyhuman tumors and this is reported to be associated with chromosomalinstability in mammary tumors (Miyoshi et al. 2001 92, 370-373).

Aurora-2 is highly expressed in multiple human tumor cell lines andlevels increase as a function of Duke's stage in primary colorectalcancers [Katayama, H. et al. (Mitotic kinase expression and colorectalcancer progression) Journal of the National Cancer Institute 91,1160-1162 (1999)]. Aurora-2 plays a role in controlling the accuratesegregation of chromosomes during mitosis. Misregulation of the cellcycle can lead to cellular proliferation and other abnormalities. Inhuman colon cancer tissue, the Aurora-2 protein has been found to beover-expressed [Bischoff et al., EMBO J., 17, 3052-3065 (1998);Schumacher et al., J. Cell Biol., 143, 1635-1646 (1998); Kimura et al.,J. Biol. Chem., 272, 13766-13771 (1997)]. Aurora-2 is over-expressed inthe majority of transformed cells. Bischoff et al. found high levels ofAurora-2 in 96% of cell lines derived from lung, colon, renal, melanomaand breast tumors (Bischoff et al. EMBO J. 1998 17, 3052-3065). Twoextensive studies show elevated Aurora-2 in 54% and 68% (Bishoff et al.EMBO J. 1998 17, 3052-3065) (Takahashi et al. 2000 Jpn J Cancer Res. 91,1007-1014) of colorectal tumours and in 94% of invasive ductadenocarcinomas of the breast (Tanaka et al. 1999 59, 2041-2044).

Aurora-1 expression is elevated in cell lines derived from tumors of thecolon, breast, lung, melanoma, kidney, ovary, pancreas, CNS, gastrictract and leukemias (Tatsuka et al. 1998 58, 4811-4816).

High levels of Aurora-3 have been detected in several tumour cell lines,although it is restricted to testis in normal tissues (Kimura et al.1999 274, 7334-7340). Over-expression of Aurora-3 in a high percentage(c. 50%) of colorectal cancers has also been documented (Takahashi etal. 2000 Jpn J Cancer Res. 91, 1007-1014). In contrast, the Aurorafamily is expressed at a low level in the majority of normal tissues,the exceptions being tissues with a high proportion of dividing cellssuch as the thymus and testis (Bischoff et al. EMBO J. 1998 17,3052-3065).

For further review of the role Aurora kinases play in proliferativedisorders, see Bischoff, J. R. & Plowman, G. D. (The Aurora/Ipl1p kinasefamily:regulators of chromosome segregation and cytokinesis) Trends inCell Biology 9, 454-459 (1999); Giet, R. and Prigent, C.(Aurora/Ipl1p-related kinases, a new oncogenic family of mitoticserine-threonine kinases) Journal of Cell Science 112, 3591-3601 (1999);Nigg, E. A. (Mitotic kinases as regulators of cell division and itscheckpoints) Nat. Rev. Mol. Cell Biol. 2, 21-32 (2001); Adams, R. R,Carmena, M., and Earnshaw, W. C. (Chromosomal passengers and the(aurora) ABCs of mitosis) Trends in Cell Biology 11, 49-54 (2001); andDutertre, S., Descamps, S., & Prigent, P. (On the role of aurora-A incentrosome function) Oncogene 21, 6175-6183 (2002).

Cyclin-dependent kinases (CDKs) are serine/threonine protein kinasesconsisting of a β-sheet rich amino-terminal lobe and a largercarboxy-terminal lobe that is largely α-helical. The CDKs display the 11subdomains shared by all protein kinases and range in molecular massfrom 33 to 44 kD. This family of kinases, which includes CDK1, CKD2,CDK4, and CDK6, requires phosphorylation at the residue corresponding toCDK2 Thr160 in order to be fully active [Meijer, L., Drug ResistanceUpdates 2000, 3, 83-88].

Each CDK complex is formed from a regulatory cyclin subunit (e.g.,cyclin A, B1, B2, D1, D2, D3, and E) and a catalytic kinase subunit(e.g., CDK1, CDK2, CDK4, CDK5, and CDK6). Each different kinase/cyclinpair functions to regulate the different and specific phases of the cellcycle known as the G1, S, G2, and M phases [Nigg, E., Nature Reviews2001, 2, 21-32; Flatt, P., Pietenpol, J., Drug Metabolism Reviews 2000,32, 283-305].

The CDKs have been implicated in cell proliferation disorders,particularly in cancer. Cell proliferation is a result of the direct orindirect deregulation of the cell division cycle and the CDKs play acritical role in the regulation of the various phases of this cycle. Forexample, the over-expression of cyclin D1 is commonly associated withnumerous human cancers including breast, colon, hepatocellularcarcinomas and gliomas [Flatt, P., Pietenpol, J., Drug MetabolismReviews 2000, 32, 283-305]. The CDK2/cyclin E complex plays a key rolein the progression from the early G₁ to S phases of the cell cycle andthe overexpression of cyclin E has been associated with various solidtumors. Therefore, inhibitors of cyclins D1, E, or their associated CDKsare useful targets for cancer therapy [Kaubisch, A., Schwartz, G., TheCancer Journal 2000, 6, 192-212].

CDKs, especially CDK2, also play a role in apoptosis and T-celldevelopment. CDK2 has been identified as a key regulator of thymocyteapoptosis [Williams, O., et al, European Journal of Immunology 2000,709-713]. Stimulation of CDK2 kinase activity is associated with theprogression of apoptosis in thymocytes, in response to specific stimuli.Inhibition of CDK2 kinase activity blocks this apoptosis resulting inthe protection of thymocytes.

In addition to regulating the cell cycle and apoptosis, the CDKs aredirectly involved in the process of transcription. Numerous virusesrequire CDKs for their replication process. Examples where CDKinhibitors restrain viral replication include human cytomegalovirus,herpes virus, and varicella-zoster virus [Meijer, L., Drug ResistanceUpdates 2000, 3, 83-88].

Inhibition of CDK is also useful for the treatment of neurodegenerativedisorders such as Alzheimer's disease. The appearance of Paired HelicalFilaments (PHF), associated with Alzheimer's disease, is caused by thehyperphosphorylation of Tau protein by CDK5/p25 [Meijer, L., DrugResistance Updates, 2000 3, 83-88].

Glycogen synthase kinase-3 (GSK-3) is a serine/threonine protein kinasecomprised of α and β isoforms that are each encoded by distinct genes[Coghlan et al., Chemistry & Biology 2000, 7, 793-803; and Kim andKimmel, Curr. Opinion Genetics Dev., 2000 10, 508-514]. GSK-3 has beenimplicated in various diseases including diabetes, Alzheimer's disease,CNS disorders such as manic depressive disorder and neurodegenerativediseases, and cardiomyocyte hypertrophy [PCT Application Nos.: WO99/65897 and WO 00/38675; and Haq et al., J. Cell Biol. 2000, 151,117-130]. These diseases are associated with the abnormal operation ofcertain cell signaling pathways in which GSK-3 plays a role. GSK-3 hasbeen found to phosphorylate and modulate the activity of a number ofregulatory proteins. These proteins include glycogen synthase, which isthe rate limiting enzyme necessary for glycogen synthesis, themicrotubule associated protein Tau, the gene transcription factorβ-catenin, the translation initiation factor e1F2B, as well as ATPcitrate lyase, axin, heat shock factor-1, c-Jun, c-myc, c-myb, CREB, andCEPBα. These diverse protein targets implicate GSK-3 in many aspects ofcellular metabolism, proliferation, differentiation, and development.

In a GSK-3 mediated pathway that is relevant for the treatment of typeII diabetes, insulin-induced signaling leads to cellular glucose uptakeand glycogen synthesis. Along this pathway, GSK-3 is a negativeregulator of the insulin-induced signal. Normally, the presence ofinsulin causes inhibition of GSK-3 mediated phosphorylation anddeactivation of glycogen synthase. The inhibition of GSK-3 leads toincreased glycogen synthesis and glucose uptake [Klein et al., PNAS1996, 93, 8455-8459; Cross et al., Biochem. J. 1994, 303, 21-26); Cohen,Biochem. Soc. Trans. 1993, 21, 555-567; and Massillon et al., Biochem J.1994, 299, 123-128]. However, in a diabetic patient, where the insulinresponse is impaired, glycogen synthesis and glucose uptake fail toincrease despite the presence of relatively high blood levels ofinsulin. This leads to abnormally high blood levels of glucose withacute and long-term effects that may ultimately result in cardiovasculardisease, renal failure and blindness. In such patients, the normalinsulin-induced inhibition of GSK-3 fails to occur. It has also beenreported that in patients with type II diabetes, GSK-3 is overexpressed[see, PCT Application: WO 00/38675]. Therapeutic inhibitors of GSK-3 aretherefore potentially useful for treating diabetic patients sufferingfrom an impaired response to insulin.

GSK-3 activity is associated with Alzheimer's disease. This disease ischaracterized by the well-known β-amyloid peptide and the formation ofintracellular neurofibrillary tangles. The neurofibrillary tanglescontain hyperphosphorylated Tau protein, in which Tau is phosphorylatedon abnormal sites. GSK-3 is known to phosphorylate these abnormal sitesin cell and animal models. Furthermore, inhibition of GSK-3 has beenshown to prevent hyperphosphorylation of Tau in cells [Lovestone et al.,Current Biology 1994, 4, 1077-86; and Brownlees et al., Neuroreport1997, 8, 3251-55]. Therefore, GSK-3 activity promotes generation of theneurofibrillary tangles and the progression of Alzheimer's disease.

Another substrate of GSK-3 is β-catenin, which is degradated afterphosphorylation by GSK-3. Reduced levels of β-catenin have been reportedin schizophrenic patients and have also been associated with otherdiseases related to increase in neuronal cell death [Zhong et al.,Nature 1998, 395, 698-702; Takashima et al., PNAS 1993, 90, 7789-93; andPei et al., J. Neuropathol. Exp 1997, 56, 70-78].

GSK-3 activity is associated with stroke [Wang et al., Brain Res 2000,859, 381-5; Sasaki et al., Neurol Res 2001, 23, 588-92; Hashimoto etal., J. Biol. Chem 2002, 277, 32985-32991].

The Janus kinases (JAK) are a family of tyrosine kinases consisting ofJAK1, JAK2, JAK3 and TYK2. The JAKs play a critical role in cytokinesignaling. The down-stream substrates of the JAK family of kinasesinclude the signal transducer and activator of transcription (STAT)proteins. JAK/STAT signaling has been implicated in the mediation ofmany abnormal immune responses such as allergies, asthma, autoimmunediseases such as transplant rejection, rheumatoid arthritis, amyotrophiclateral sclerosis and multiple sclerosis as well as in solid andhematologic malignancies such as leukemias and lymphomas. Thepharmaceutical intervention in the JAK/STAT pathway has been reviewed[Frank Mol. Med. 1999, 5, 432-456 and Seidel et al., Oncogene 2000, 19,2645-2656].

JAK1, JAK2, and TYK2 are ubiquitously expressed, while JAK3 ispredominantly expressed in hematopoietic cells. JAK3 binds exclusivelyto the common cytokine receptor gamma chain (γ_(c)) and is activated byIL-2, IL-4, IL-7, IL-9, and IL-15. The proliferation and survival ofmurine mast cells induced by IL-4 and IL-9 have, in fact, been shown tobe dependent on JAK3- and γ_(c)-signaling [Suzuki et al., Blood 2000,96, 2172-2180].

Cross-linking of the high-affinity immunoglobulin (Ig) E receptors ofsensitized mast cells leads to a release of proinflammatory mediators,including a number of vasoactive cytokines resulting in acute allergic,or immediate (type I) hypersensitivity reactions [Gordon et al., Nature1990, 346, 274-276 and Galli, N. Engl. J. Med. 1993, 328, 257-265]. Acrucial role for JAK3 in IgE receptor-mediated mast cell responses invitro and in vivo has been established [Malaviya et al., Biochem.Biophys. Res. Commun. 1999, 257, 807-813]. In addition, the preventionof type I hypersensitivity reactions, including anaphylaxis, mediated bymast cell-activation through inhibition of JAK3 has also been reported[Malaviya et al., J. Biol. Chem. 1999 274, 27028-27038]. Targeting mastcells with JAK3 inhibitors modulated mast cell degranulation in vitroand prevented IgE receptor/antigen-mediated anaphylactic reactions invivo.

A recent study described the successful targeting of JAK3 forimmunosuppression and allograft acceptance. The study demonstrated adose-dependent survival of Buffalo heart allograft in Wistar Furthrecipients upon administration of inhibitors of JAK3 indicating thepossibility of regulating unwanted immune responses in graft versus hostdisease [Kirken, Transpl. Proc. 2001, 33, 3268-3270].

IL-4-mediated STAT-phosphorylation has been implicated as the mechanisminvolved in early and late stages of rheumatoid arthritis (RA).Up-regulation of proinflammatory cytokines in RA synovium and synovialfluid is a characteristic of the disease. It has been demonstrated thatIL-4-mediated activation of IL-4/STAT pathway is mediated through theJanus Kinases (JAK 1 & 3) and that IL-4-associated JAK kinases areexpressed in the RA synovium [Muller-Ladner et al., J. Immunol. 2000,164, 3894-3901].

Familial amyotrophic lateral sclerosis (FALS) is a fatalneurodegenerative disorder affecting about 10% of ALS patients. Thesurvival rates of FALS mice were increased upon treatment with a JAK3specific inhibitor. This suggested that JAK3 plays a role in FALS [Trieuet al., Biochem. Biophys. Res. Commun. 2000, 267, 22-25].

Signal transducer and activator of transcription (STAT) proteins areactivated by, among others, the JAK family kinases. Results from arecent study suggested the possibility of intervention in the JAK/STATsignaling pathway by targeting JAK family kinases with specificinhibitors for the treatment of leukemia [Sudbeck et al., Clin. CancerRes. 1999, 5, 1569-1582]. JAK3 specific compounds were shown to inhibitthe clonogenic growth of JAK3-expressing cell lines DAUDI, RAMOS,LC1-19, NALM-6, MOLT-3 and HL-60.

In animal models, TEL/JAK2 fusion proteins have inducedmyeloproliferative disorders and in hematopoietic cell lines, andintroduction of TEL/JAK2 resulted in activation of STAT1, STAT3, STAT5,and cytokine-independent growth [Schwaller et al., EMBO J. 1998, 17,5321-5333].

Inhibition of JAK3 and TYK2 abrogated tyrosine phosphorylation of STAT3,and inhibited cell growth of mycosis fungoides, a form of cutaneousT-cell lymphoma. These results implicated JAK family kinases in theconstitutively activated JAK/STAT pathway that is present in mycosisfungoides [Nielsen et al., Proc. Nat. Acad. Sci. U.S.A. 1997, 94,6764-6769]. Similarly, STAT3, STAT5, JAK1 and JAK2 were demonstrated tobe constitutively activated in mouse T-cell lymphoma characterizedinitially by LCK over-expression, thus further implicating the JAK/STATpathway in abnormal cell growth [Yu et al., J. Immunol. 1997, 159,5206-5210]. In addition, IL-6-mediated STAT3 activation was blocked byan inhibitor of JAK, leading to sensitization of myeloma cells toapoptosis [Catlett-Falcone et al., Immunity 1999, 10, 105-115].

Another kinase family of particular interest is the Src family ofkinases. These kinases are implicated in cancer, immune systemdysfunction and bone remodeling diseases. For general reviews, seeThomas and Brugge, Annu. Rev. Cell Dev. Biol. 1997, 13, 513; Lawrenceand Niu, Pharmacol. Ther. 1998, 77, 81; Tatosyan and Mizenina,Biochemistry (Moscow) 2000, 65, 49-58; Boschelli et al., Drugs of theFuture 2000, 25(7), 717.

Members of the Src family include the following eight kinases inmammals: Src, Fyn, Yes, Fgr, Lyn, Hck, Lck, and Blk. These arenonreceptor protein kinases that range in molecular mass from 52 to 62kD. All are characterized by a common structural organization that iscomprised of six distinct functional domains: Src homology domain 4(SH4), a unique domain, SH3 domain, SH2 domain, a catalytic domain(SH1), and a C-terminal regulatory region. Tatosyan et al. Biochemistry(Moscow) 2000, 65, 49-58.

Based on published studies, Src kinases are considered as potentialtherapeutic targets for various human diseases. Mice that are deficientin Src develop osteopetrosis, or bone build-up, because of depressedbone resorption by osteoclasts. This suggests that osteoporosisresulting from abnormally high bone resorption can be treated byinhibiting Src. Soriano et al., Cell 1992, 69, 551 and Soriano et al.,Cell 1991, 64, 693.

Suppression of arthritic bone destruction has been achieved by theoverexpression of CSK in rheumatoid synoviocytes and osteoclasts.Takayanagi et al., J. Clin. Invest. 1999, 104, 137. CSK, or C-terminalSrc kinase, phosphorylates and thereby inhibits Src catalytic activity.This implies that Src inhibition may prevent joint destruction that ischaracteristic in patients suffering from rheumatoid arthritis.Boschelli et al., Drugs of the Future 2000, 25(7), 717.

Src also plays a role in the replication of hepatitis B virus. Thevirally encoded transcription factor HBx activates Src in a steprequired for propagation of the virus. Klein et al., EMBO J. 1999, 18,5019, and Klein et al., Mol. Cell. Biol. 1997, 17, 6427.

A number of studies have linked Src expression to cancers such as colon,breast, hepatic and pancreatic cancer, certain B-cell leukemias andlymphomas. Talamonti et al., J. Clin. Invest. 1993, 91, 53; Lutz et al.,Biochem. Biophys. Res. 1998 243, 503; Rosen et al., J. Biol. Chem. 1986,261, 13754; Bolen et al., Proc. Natl. Acad. Sci. USA 1987, 84, 2251;Masaki et al., Hepatology 1998, 27, 1257; Biscardi et al., Adv. CancerRes. 1999, 76, 61; Lynch et al., Leukemia 1993, 7, 1416. Furthermore,antisense Src expressed in ovarian and colon tumor cells has been shownto inhibit tumor growth. Wiener et al., Clin. Cancer Res., 1999, 5,2164; Staley et al., Cell Growth Diff. 1997, 8, 269.

Other Src family kinases are also potential therapeutic targets. Lckplays a role in T-cell signaling. Mice that lack the Lck gene have apoor ability to develop thymocytes. The function of Lck as a positiveactivator of T-cell signaling suggests that Lck inhibitors may be usefulfor treating autoimmune disease such as rheumatoid arthritis. Molina etal., Nature, 1992, 357, 161. Hck, Fgr and Lyn have been identified asimportant mediators of integrin signaling in myeloid leukocytes. Lowellet al., J. Leukoc. Biol., 1999, 65, 313. Inhibition of these kinasemediators may therefore be useful for treating inflammation. Boschelliet al., Drugs of the Future 2000, 25(7), 717.

Syk is a tyrosine kinase that plays a critical role in FcεRI mediatedmast cell degranulation and eosinophil activation. Accordingly, Sykkinase is implicated in various allergic disorders, in particularasthma. It has been shown that Syk binds to the phosphorylated gammachain of the FcεRI receptor via N-terminal SH2 domains and is essentialfor downstream signaling [Taylor et al., Mol. Cell. Biol. 1995, 15,4149].

Inhibition of eosinophil apoptosis has been proposed as a key mechanismfor the development of blood and tissue eosinophilia in asthma. IL-5 andGM-CSF are upregulated in asthma and are proposed to cause blood andtissue eosinophilia by inhibition of eosinophil apoptosis. Inhibition ofeosinophil apoptosis has been proposed as a key mechanism for thedevelopment of blood and tissue eosinophilia in asthma. It has beenreported that Syk kinase is required for the prevention of eosinophilapoptosis by cytokines (using antisense)[Yousefi et al., J. Exp. Med.1996, 183, 1407].

The role of Syk in FcγR dependent and independent response in bonemarrow derived macrophages has been determined by using irradiated mousechimeras reconstituted with fetal liver cells from Syk −/− embryos. Sykdeficient macrophages were defective in phagocytosis induced by FcγR butshowed normal phagocytosis in response to complement [Kiefer et al.,Mol. Cell. Biol. 1998, 18, 4209]. It has also been reported thataerosolized Syk antisense suppresses Syk expression and mediator releasefrom macrophages [Stenton et al., J. Immunology 2000, 164, 3790].

The Tec family of non-receptor tyrosine kinases plays a central role insignalling through antigen-receptors such as the TCR, BCR and FCEreceptors (reviewed in Miller A, et al. Current Opinion in Immunology14; 331-340 (2002). Tec family kinases are essential for T cellactivation. Three members of the Tec family, Itk, Rlk and Tec, areactivated downstream of antigen receptor engagement in T cells andtransmit signals to downstream effectors, including PLC-γ. Deletion ofItk in mice results in reduced T cell receptor (TCR)-inducedproliferation and secretion of the cytokines IL-2, IL-4, IL-5, IL-10 andIFN-γ (Schaeffer et al, Science 284; 638-641 (1999)), Fowell et al,Immunity 11; 399-409 (1999), Schaeffer et al. Nature Immunology 2, 12;1183-1188 (2001))). The immunological symptoms of allergic asthma areattenuated in Itk−/− mice. Lung inflammation, eosinophil infiltrationand mucous production are drastically reduced in Itk−/− mice in responseto challenge with the allergen OVA (Mueller et al, Journal of Immunology170: 5056-5063 (2003)). Itk has also been implicated in atopicdermatitis. This gene has been reported to be more highly expressed inperipheral blood T cells from patients with moderate and/or severeatopic dermatitis than in controls or patients with mild atopicdermatitis (Matsumoto et al, International archives of Allergy andImmunology 129; 327-340 (2002)).

Splenocytes from Rlk−/− mice secrete half the IL-2 produced by wild typeanimals in response to TCR engagement (Schaeffer et al, Science 284;638-641 (1999)), while combined deletion of Itk and Rlk in mice leads toa profound inhibition of TCR-induced responses including proliferationand production of the cytokines IL-2, IL-4, IL-5 and IFN-γ (Schaeffer etal. Nature Immunology 2, 12; 1183-1188 (2001)), Schaeffer et al, Science284; 638-641 (1999)). Intracellular signalling following TCR engagementis effected in Itk/Rlk deficient T cells; inositol triphosphateproduction, calcium mobilization, MAP kinase activation, and activationof the transcription factors NFAT and AP-1 are all reduced (Schaeffer etal, Science 284; 638-641 (1999), Schaeffer et al. Nature Immunology 2,12; 1183-1188 (2001)).

Tec family kinases are also essential for B cell development andactivation. Patients with mutations in Btk have a profound block in Bcell development, resulting in the almost complete absence of Blymphocytes and plasma cells, severely reduced Ig levels and a profoundinhibition of humoral response to recall antigens (reviewed in Vihinenet al. Frontiers in Bioscience 5:d917-928). Mice deficient in Btk alsohave a reduced number of peripheral B cells and greatly decreased levelsof IgM and IgG3. Btk deletion in mice has a profound effect on B cellproliferation induced by anti-IgM, and inhibits immune responses tothymus-independent type II antigens (Ellmeier et al, J Exp Med192:1611-1623 (2000)).

Tec kinases also play a role in mast cell activation through thehigh-affinity IgE receptor (FcεRI). Itk and Btk are expressed in mastcells and are activated by FcεRI cross-linking (Kawakami et al, Journalof Immunology; 3556-3562 (1995)). Btk deficient murine mast cells havereduced degranulation and decreased production of proinflammatorycytokines following FcεRI cross-linking (Kawakami et al. Journal ofleukocyte biology 65:286-290). Btk deficiency also results in a decreaseof macrophage effector functions (Mukhopadhyay et al, Journal ofImmunology; 168, 2914-2921 (2002)).

Accordingly, there is a great need to develop compounds useful asinhibitors of protein kinases. In particular, it would be desirable todevelop compounds that are useful as inhibitors of GSK-3, SYK, Aurora-2,CDK-2, JAK-3, LCK, SRC, and Tec family (e.g., Tec, Btk, Itk/Emt/Tsk,Bmx, Txk/Rlk) protein kinases, particularly given the inadequatetreatments currently available for the majority of the disordersimplicated in their activation.

SUMMARY OF THE INVENTION

It has now been found that compounds of this invention, andpharmaceutically acceptable compositions thereof, are effective asinhibitors of protein kinases. In certain embodiments, these compoundsare effective as inhibitors of GSK-3, SYK, Aurora-2, CDK-2, JAK-3, LCK,SRC, and Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) proteinkinases. These compounds have the general formula I:

-   -   or a pharmaceutically acceptable salt thereof, wherein R¹, R²,        Cy¹, T, n and X are as defined below.

These compounds and pharmaceutically acceptable compositions thereof areuseful for treating or preventing a variety of diseases, disorders orconditions, including, but not limited to, heart disease, diabetes,Alzheimer's disease, immunodeficiency disorders, inflammatory diseases,allergic diseases, autoimmune diseases, destructive bone disorders suchas osteoporosis, proliferative disorders, infectious diseases,immunologically-mediated diseases, neurodegenerative or neurologicaldisorders, or viral diseases. The compositions are also useful inmethods for preventing cell death and hyperplasia and therefore may beused to treat or prevent reperfusion/ischemia in stroke, heart attacks,and organ hypoxia. The compositions are also useful in methods forpreventing thrombin-induced platelet aggregation.

The compounds provided by this invention are also useful for the studyof kinases in biological and pathological phenomena; the study ofintracellular signal transduction pathways mediated by such kinases; andthe comparative evaluation of new kinase inhibitors.

DETAILED DESCRIPTION OF THE INVENTION

I. General Description of Compounds of the Invention:

The present invention relates to a compound of formula I:

-   -   or a pharmaceutically acceptable salt thereof, wherein:    -   X is an optionally substituted C₁-C₃ alkylidene chain wherein        one or two non-adjacent methylene units are independently        optionally replaced by CO, CONR′, NR′CO, SO, SO₂, NR′SO₂,        SO₂NR′, O, S, or NR′;

R¹ and R², taken together with the carbon atoms to which they are boundform an optionally substituted 5- or 6-membered aryl or heteroaryl ringhaving 0-3 heteroatoms independently selected from nitrogen, oxygen, orsulfur,

-   -   wherein the ring formed by R¹ and R² taken together, and the        C₁-C₃ alkylidene chain of X, are each optionally and        independently substituted at one or more carbon atoms with y        occurrences of —WR^(y), wherein y is 0-5; and wherein one or        more substitutable nitrogen atoms of the ring formed by R¹ and        R² taken together are optionally substituted with —R³;

each occurrence of W is independently a bond or is a C₁-C₆ alkylidenechain wherein up to two non-adjacent methylene units of W areindependently optionally replaced by CO, CO₂, COCO, CONR, OCONR, NRNR,NRNRCO, NRCO, NRCO₂, NRCONR, SO, SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, orNR; and

each occurrence of R^(y) is independently selected from R′, halogen,NO₂, or CN, or —WR^(y) is ═O, ═S, or ═NR′;

T is CHR′, CH₂CH(R′), —S(═O)₂, or C(═O);

n is 0 or 1;

Cy¹ is a 3-7 membered saturated, partially unsaturated, or fullyunsaturated monocyclic ring having 0-3 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur, or an 8-10 memberedsaturated, partially unsaturated, or fully unsaturated bicyclic ringsystem having 0-5 heteroatoms independently selected from nitrogen,oxygen, or sulfur, wherein Cy¹ is optionally substituted at one or morecarbon atoms with x independent occurrences of -QR^(x); wherein x is0-5; and at one or more substitutable nitrogen atoms with —R⁴; wherein

Q is a bond or is a C₁-C₆ alkylidene chain wherein up to twonon-adjacent methylene units of Q are independently optionally replacedby CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO, NRCO₂, NRCONR, SO,SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR; and each occurrence of R^(x) isindependently selected from R′, halogen, NO₂, or CN, or-QR^(x) is ═O,═S, or ═NR′;

each occurrence of R³ and R⁴ is independently R′, —COR′, —CO₂(C₁₋₆aliphatic), —CON(R′)₂, or —SO₂R′;

each occurrence of R is independently selected from hydrogen or anoptionally substituted C₁₋₆ aliphatic group; and

each occurrence of R′ is independently selected from hydrogen or anoptionally substituted group selected from C₁₋₈ aliphatic, C₆₋₁₀ aryl, aheteroaryl ring having 5-10 ring atoms, or a heterocyclyl ring having3-10 ring atoms, or wherein R and R′ taken together with the atom(s) towhich they are bound, or two occurrences of R′ taken together with theatom(s) to which they are bound, form an optionally substituted 3-8membered cycloalkyl, heterocyclyl, aryl, or heteroaryl ring having 0-3heteroatoms independently selected from nitrogen, oxygen, or sulfur; and

the dashed bond represents a single or double bond, as valency permits;

provided that:

a) when n is 0, Cy¹ is an optionally substituted aryl or heteroarylgroup, and X is —(C(R)₂)₂—, —CH₂—O—, —O—CH₂—, —S—CH₂—, CH₂—S—,—CH₂—N(R)—, —N(R)—CH₂—, —CH₂SO—, —SOCH₂—, wherein R is hydrogen orC₁₋₄alkyl, then R¹ and R², taken together with the carbon atoms to whichthey are bound do not form a phenyl ring optionally substituted withC₁₋₄alkyl, C₁₋₄alkoxy, halo, CONR₂, CO₂R, fused phenyl, NO₂, ortrifluoromethyl;

b) when n is 0, Cy¹ is 3,4,5-trimethoxyphenyl, and when X is —(CH₂)₂—,then R′ and R², taken together with the carbon atoms to which they arebound, do not form an unsubstituted thieno ring; and

c) when X is —(CH₂)₂—, n is 0 and Cy¹ is a phenyl group bearing one ormore of the following substitutents: —(CH₂)₂OH, —O(CH₂)₂OH, —(CH₂)₂NMe₂,—O(CH₂)₂NHEt, —O(CH_(—2))₂NEt₂, —OMe and —O(CH₂)₂OS(O)₂-p-Tol, then R¹and R², taken together with the carbon atoms to which they are bound, donot form a methoxy-substituted pyridyl ring.

In certain other embodiments for compounds described directly above:

a) when n is 0, Cy¹ is an optionally substituted aryl or heteroarylgroup, and X is —(C(R)₂)₂—, —CH₂—O—, —O—CH₂—, —S—C(R)₂—, —C(R)₂—S—,—CH₂—N(R)—, —N(R)—CH₂—, —CH₂SO—, —SOCH₂—, —N(R)SO₂—, C═O, —CH₂—, C═C,—N(R)N(R)—, or N═N, wherein R is hydrogen or C₁₋₄alkyl, then R¹ and R²,taken together with the carbon atoms to which they are bound do not forma phenyl ring optionally substituted with —C₁₋₄alkyl, —C₁₋₄alkoxy,-halo, —CONR₂, —CO₂R, fused phenyl, —NO₂, —NR₂, —N(CO)R, —NSO₂R,—N(CO)N(R)₂, —SCH₃, —SR, —CH₂OH, —OH, —C(O)H, —OCH₂Ph, —N(CO)OR,2-furyl, or trifluoromethyl;

b) when n is 0 and Cy¹ is 3,4,5-trimethoxyphenyl then

-   -   i) when X is —(CH₂)₂—, then R¹ and R², taken together with the        carbon atoms to which they are bound, do not form an        unsubstituted thieno ring; and    -   ii) when X is —S—CR₂—, then R¹ and R², taken together with the        carbon atoms to which they are bound, do not form an        unsubstituted or a chloro-substituted phenyl ring (or in certain        embodiments, an unsubstituted or a substituted phenyl); and    -   iii) when X is —CH₂CH₂CH₂—, then R¹ and R², taken together with        the carbon atoms to which they are bound, do not form an        unsubstituted phenyl ring (or in certain embodiments, a        substituted or unsubstituted phenyl); and

c) when X is —(CH₂)₂—, n is 0 and Cy¹ is a phenyl group bearing one ormore of the following substitutents: —(CH₂)₂OH, —O(CH₂)₂OH, —(CH₂)₂NMe₂,—O(CH₂)₂NHEt, —O(CH₂)₂NEt₂, —OMe and —O(CH₂)₂OS(O)₂-p-Tol, then R¹ andR², taken together with the carbon atoms to which they are bound, do notform a methoxy-substituted (or in certain embodiments, a substituted orunsubstituted) pyridyl ring.

d) when n is 1, T is CH₂, and X is —OC(H)(OH)—, Cy is not optionallysubstituted pyridyl.

In certain other embodiments for compounds described above:

-   -   a. when n is 0, Cy¹ is an optionally substituted phenyl,        benzotriazolyl, benzothiazolyl, indolyl, or pyridyl group, and X        is a C₁₋₂alkylene group optionally substituted with one or more        halogen atoms or C₁₋₃alkyl groups, then R¹ and R², taken        together with the carbon atoms to which they are bound do not        form a pyridyl, or thienyl group, when the pyridyl or thienyl        group is unsubstituted or is substituted with a substituent        other than a 3-7 membered saturated, partially unsaturated, or        fully unsaturated monocyclic ring having 0-3 heteroatoms        independently selected from nitrogen, oxygen, or sulfur, or an        8-10 membered saturated, partially unsaturated, or fully        unsaturated bicyclic ring system having 0-5 heteroatoms        independently selected from nitrogen, oxygen, or sulfur;    -   b. when n is 0, Cy¹ is an optionally substituted C₆-C₁₂ mono- or        bicyclic aromatic group, and X is a C₁₋₂alkylene group        optionally substituted with one or more halogen atoms or        C₁₋₃alkyl groups, then R¹ and R², taken together with the carbon        atoms to which they are bound do not form a phenyl group, when        the phenyl group is unsubstituted or is substituted with a        substituent other than a 3-7 membered saturated, partially        unsaturated, or fully unsaturated monocyclic ring having 0-3        heteroatoms independently selected from nitrogen, oxygen, or        sulfur, or an 8-10 membered saturated, partially unsaturated, or        fully unsaturated bicyclic ring system having 0-5 heteroatoms        independently selected from nitrogen, oxygen, or sulfur;    -   c. when n is 0; Cy¹ is an aryl or heteroaryl group substituted        with at least one optionally substituted cycloaliphatic or        heterocycloaliphatic group, wherein the cycloaliphatic or        heterocycloaliphatic groups are attached directly to Cy¹ or are        attached to Cy¹ through an alkylidene chain; and X is an        optionally substituted C₁₋₂alkylene group; then R¹ and R², taken        together with the carbon atoms to which they are bound do not        form an optionally substituted phenyl group.

In certain other embodiments, this invention provides a compound offormula I, wherein the optionally substituted ring containing X is anaromatic ring (either 5- or 6-membered aromatic ring) and R¹ and R²,taken together with the carbon atoms to which they are bound form anoptionally substituted 5- or 6-membered heteroaryl ring having 0-3heteroatoms independently selected from nitrogen, oxygen, or sulfur. Inpreferred forms of this embodiment, the heteroaryl ring is an optionallysubstituted thienyl or thiazolyl ring. It should be understood that theoptional substituents and the other variables in this embodiment are asdefined above or in any embodiment herein.

2. Compounds and Definitions:

Compounds of this invention include those described generally above, andare further illustrated by the classes, subclasses, and speciesdisclosed herein. As used herein, the following definitions shall applyunless otherwise indicated. For purposes of this invention, the chemicalelements are identified in accordance with the Periodic Table of theElements, CAS version, Handbook of Chemistry and Physics, 75^(th) Ed.Additionally, general principles of organic chemistry are described in“Organic Chemistry”, Thomas Sorrell, University Science Books,Sausalito: 1999, and “March's Advanced Organic Chemistry”, 5^(th) Ed.:Smith, M. B. and March, J., John Wiley & Sons, New York: 2001, theentire contents of which are hereby incorporated by reference.

As described herein, compounds of the invention may optionally besubstituted with one or more substituents, such as are illustratedgenerally above, or as exemplified by particular classes, subclasses,and species of the invention. It will be appreciated that the phrase“optionally substituted” is used interchangeably with the phrase“substituted or unsubstituted.” In general, the term “substituted”,whether preceded by the term “optionally” or not, refers to thereplacement of hydrogen radicals in a given structure with the radicalof a specified substituent. Unless otherwise indicated, an optionallysubstituted group may have a substituent at each substitutable positionof the group, and when more than one position in any given structure maybe substituted with more than one substituent selected from a specifiedgroup, the substituent may be either the same or different at everyposition. Combinations of substituents envisioned by this invention arepreferably those that result in the formation of stable or chemicallyfeasible compounds. The term “stable”, as used herein, refers tocompounds that are not substantially altered when subjected toconditions to allow for their production, detection, and preferablytheir recovery, purification, and use for one or more of the purposesdisclosed herein. In some embodiments, a stable compound or chemicallyfeasible compound is one that is not substantially altered when kept ata temperature of 40° C. or less, in the absence of moisture or otherchemically reactive conditions, for at least a week.

The term “aliphatic” or “aliphatic group”, as used herein, means astraight-chain (i.e., unbranched) or branched, substituted orunsubstituted hydrocarbon chain that is completely saturated or thatcontains one or more units of unsaturation, or a monocyclic hydrocarbonor bicyclic hydrocarbon that is completely saturated or that containsone or more units of unsaturation, but which is not aromatic (alsoreferred to herein as “carbocycle” “cycloaliphatic” or “cycloalkyl”),that has a single point of attachment to the rest of the molecule.Unless otherwise specified, aliphatic groups contain 1-20 aliphaticcarbon atoms. In some embodiments, aliphatic groups contain 1-10aliphatic carbon atoms. In other embodiments, aliphatic groups contain1-8 aliphatic carbon atoms. In still other embodiments, aliphatic groupscontain 1-6 aliphatic carbon atoms, and in yet other embodimentsaliphatic groups contain 1-4 aliphatic carbon atoms. In someembodiments, “cycloaliphatic” (or “carbocycle” or “cycloalkyl”) refersto a monocyclic C₃-C₈ hydrocarbon or bicyclic C₈-C₁₂ hydrocarbon that iscompletely saturated or that contains one or more units of unsaturation,but which is not aromatic, that has a single point of attachment to therest of the molecule wherein any individual ring in said bicyclic ringsystem has 3-7 members. Suitable aliphatic groups include, but are notlimited to, linear or branched, substituted or unsubstituted alkyl,alkenyl, alkynyl groups and hybrids thereof such as (cycloalkyl)alkyl,(cycloalkenyl)alkyl or (cycloalkyl)alkenyl.

The term “alkylene” or “alkylene group”, as used herein, refers to ahydrocarbon group that is completely saturated.

The term “heteroaliphatic”, as used herein, means aliphatic groupswherein one or two carbon atoms are independently replaced by one ormore of oxygen, sulfur, nitrogen, phosphorus, or silicon.Heteroaliphatic groups may be substituted or unsubstituted, branched orunbranched, cyclic or acyclic, and include “heterocycle”,“heterocyclyl”, “heterocycloaliphatic”, or “heterocyclic” groups.

The term “heterocycle”, “heterocyclyl”, “heterocycloaliphatic”, or“heterocyclic” as used herein means non-aromatic, monocyclic, bicyclic,or tricyclic ring systems in which one or more ring members are anindependently selected heteroatom. In some embodiments, the“heterocycle”, “heterocyclyl”, “heterocycloaliphatic”, or “heterocyclic”group has three to fourteen ring members in which one or more ringmembers is a heteroatom independently selected from oxygen, sulfur,nitrogen, or phosphorus, and each ring in the system contains 3 to 7ring members.

The term “heteroatom” means one or more of oxygen, sulfur, nitrogen,phosphorus, or silicon (including, any oxidized form of nitrogen,sulfur, phosphorus, or silicon; the quaternized form of any basicnitrogen or; a substitutable nitrogen of a heterocyclic ring, forexample N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) orNR⁺ (as in N-substituted pyrrolidinyl)).

The term “unsaturated”, as used herein, means that a moiety has one ormore units of unsaturation.

The term “alkoxy”, or “thioalkyl”, as used herein, refers to an alkylgroup, as previously defined, attached to the principal carbon chainthrough an oxygen (“alkoxy”) or sulfur (“thioalkyl”) atom.

The terms “haloalkyl”, “haloalkenyl” and “haloalkoxy” means alkyl,alkenyl or alkoxy, as the case may be, substituted with one or morehalogen atoms. The term “halogen” means F, Cl, Br, or I. It should beunderstood that 1) an aliphatic group (e.g., alkyl) substituted with ahalogen could also be referred to as a haloaliphatic (e.g., haloalkyl,in the case of an halo-substituted alkyl group); and 2) thatperfluorinated groups are within the scope of this invention.

The term “aryl” used alone or as part of a larger moiety as in“aralkyl”, “aralkoxy”, or “aryloxyalkyl”, refers to monocyclic,bicyclic, and tricyclic ring systems having a total of five to fourteenring members, wherein at least one ring in the system is aromatic andwherein each ring in the system contains 3 to 7 ring members. The term“aryl” may be used interchangeably with the term “aryl ring”.

The term “heteroaryl”, used alone or as part of a larger moiety as in“heteroaralkyl” or “heteroarylalkoxy”, refers to monocyclic, bicyclic,and tricyclic ring systems having a total of five to fourteen ringmembers, wherein at least one ring in the system is aromatic, at leastone ring in the system contains one or more heteroatoms, and whereineach ring in the system contains 3 to 7 ring members. The term“heteroaryl” may be used interchangeably with the term “heteroaryl ring”or the term “heteroaromatic”.

An aryl (including aralkyl, aralkoxy, aryloxyalkyl and the like) orheteroaryl (including heteroaralkyl and heteroarylalkoxy and the like)group may contain one or more substituents. Suitable substituents on theunsaturated carbon atom of an aryl or heteroaryl group are selected fromhalogen; —R^(o); —OR^(o); —SR^(o); 1,2-methylenedioxy;1,2-ethylenedioxy; phenyl (Ph) optionally substituted with R^(o); —O(Ph)optionally substituted with R^(o); —(CH₂)₁₋₂(Ph), optionally substitutedwith R^(o); —CH═CH(Ph), optionally substituted with R^(o); —NO₂; —CN;—N(R^(o))₂; —NR^(o)C(O)R^(o); —NR^(o)C(S)R^(o); —NR^(o)C(O)N(R^(o))₂;—NR^(o)C(S)N(R^(o))₂; —NR^(o)CO₂R^(o); —NR^(o)NR^(o)C(O)R^(o);—NR^(o)NR^(o)C(O)N(R^(o))₂; —NR^(o)NR_(o)CO₂R^(o); —C(O)C(O)R^(o);—C(O)CH₂C(O)R^(o); —CO₂R^(o); —C(O)R^(o); —C(S)R^(o); —C(O)N(R^(o))₂;—C(S)N(R^(o))₂; —OC(O)N(R^(o))₂; —OC(O)R^(o); —C(O)N(OR^(o))R^(o);—C(NOR^(o))R^(o); —S(O)₂R^(o); —S(O)₃R^(o); —SO₂N(R^(o))₂; —S(O)R^(o);—NR^(o)SO₂N(R^(o))₂; —NR^(o)SO₂R^(o); —N(OR^(o))R^(o);—C(═NH)—N(R^(o))₂; or —(CH₂)₀₋₂NHC(O)R^(o) wherein each independentoccurrence of R^(o) is selected from hydrogen, optionally substitutedC₁₋₆ aliphatic, an unsubstituted 5-6 membered heteroaryl or heterocyclicring (provided that a nitrogen atom in the heterocyclic ring isoptionally substituted with —R⁺ or —C(O)R⁺, wherein R⁺ is (C₁₋₆alkyl),preferably (C₁₋₄alkyl)), phenyl, —O(Ph), or —CH₂(Ph), or,notwithstanding the definition above, two independent occurrences ofR^(o), on the same substituent or different substituents, taken togetherwith the atom(s) to which each R^(o) group is bound, form a 5-8-memberedheterocyclyl, aryl, or heteroaryl ring or a 3-8-membered cycloalkyl ringhaving 0-3 heteroatoms independently selected from nitrogen, oxygen, orsulfur. Optional substituents on the aliphatic group of R^(o) areselected from NH₂, NH(C₁ aliphatic), N(C₁ aliphatic)₂, halogen,C₁₋₄aliphatic, OH, O(C₁₋₄aliphatic), NO₂, CN, CO₂H, CO₂(C₁₋₄aliphatic),O(haloC₁₋₄ aliphatic), or haloC₁₋₄aliphatic, wherein each of theforegoing C₁₋₄aliphatic groups of R^(o) is unsubstituted.

An aliphatic or heteroaliphatic group, or a non-aromatic heterocyclicring may contain one or more substituents. Suitable substituents on thesaturated carbon of an aliphatic or heteroaliphatic group, or of anon-aromatic heterocyclic ring are selected from those listed above forthe unsaturated carbon of an aryl or heteroaryl group and additionallyinclude the following: ═O, ═S, ═NNHR*, ═NN(R*)₂, ═NNHC(O)R*,═NNHCO₂(alkyl), ═NNHSO₂(alkyl), or ═NR*, where each R* is independentlyselected from hydrogen or an optionally substituted C₁₋₆ aliphatic.Optional substituents on the aliphatic group of R* are selected fromNH₂, NH(C₁₋₄ aliphatic), N(C₁₋₄ aliphatic)₂, halogen, C₁₋₄ aliphatic,OH, O(C₁₋₄ aliphatic), NO₂, CN, CO₂H, CO₂(C₁₋₄ aliphatic), O(halo C₁₋₄aliphatic), or halo(C₁₋₄ aliphatic), wherein each of the foregoingC₁₋₄aliphatic groups of R* is unsubstituted.

Optional substituents on the nitrogen of a non-aromatic heterocyclicring are selected from —R⁺, —N(R⁺)₂, —C(O)R⁺, —CO₂R⁺, —C(O)C(O)R⁺,—C(O)CH₂C(O)R⁺, —SO₂R⁺, —SO₂N(R⁺)₂, —C(═S)N(R⁺)₂, —C(═NH)—N(R⁺)₂, or—NR⁺SO₂R⁺; wherein R⁺ is hydrogen, an optionally substituted C₁₋₆aliphatic, optionally substituted phenyl, optionally substituted —O(Ph),optionally substituted —CH₂(Ph), optionally substituted —(CH₂)₁₋₂(Ph);optionally substituted —CH═CH(Ph); or an unsubstituted 5-6 memberedheteroaryl or heterocyclic ring having one to four heteroatomsindependently selected from oxygen, nitrogen, or sulfur, or,notwithstanding the definition above, two independent occurrences of R⁺,on the same substituent or different substituents, taken together withthe atom(s) to which each R⁺ group is bound, form a 5-8-memberedheterocyclyl, aryl, or heteroaryl ring or a 3-8-membered cycloalkyl ringhaving 0-3 heteroatoms independently selected from nitrogen, oxygen, orsulfur. Optional substituents on the aliphatic group or the phenyl ringof R⁺ are selected from NH₂, NH(C₁₋₄ aliphatic), N(C₁₋₄ aliphatic)₂,halogen, C₁₋₄ aliphatic, OH, O(C₁₋₄ aliphatic), NO₂, CN, CO₂H, CO₂(C₁₋₄aliphatic), O(halo C₁₋₄ aliphatic), or halo(C₁₋₄ aliphatic), whereineach of the foregoing C₁₋₄aliphatic groups of R⁺ is unsubstituted.

The term “alkylidene chain” refers to a straight or branched carbonchain that may be fully saturated or have one or more units ofunsaturation and has two points of attachment to the rest of themolecule, wherein one or more methylene units may optionally andindependently be replaced with a group including, but not limited to,CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO, NRCO₂, NRCONR, SO, SO₂,NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR.

As detailed above, in some embodiments, two independent occurrences ofR^(o) (or R⁺, or any other variable similarly defined herein), are takentogether with the atom(s) to which each variable is bound to form a5-8-membered heterocyclyl, aryl, or heteroaryl ring or a 3-8-memberedcycloalkyl ring having 0-3 heteroatoms independently selected fromnitrogen, oxygen, or sulfur. Exemplary rings that are formed when twoindependent occurrences of R^(o) (or R⁺, or any other variable similarlydefined herein) are taken together with the atom(s) to which eachvariable is bound include, but are not limited to the following: a) twoindependent occurrences of R^(o) (or R⁺, or any other variable similarlydefined herein) that are bound to the same atom and are taken togetherwith that atom to form a ring, for example, N(R^(o))₂, where bothoccurrences of R^(o) are taken together with the nitrogen atom to form apiperidin-1-yl, piperazin-1-yl, or morpholin-4-yl group; and b) twoindependent occurrences of R^(o) (or R⁺, or any other variable similarlydefined herein) that are bound to different atoms and are taken togetherwith both of those atoms to form a ring, for example where a phenylgroup is substituted with two occurrences of OR^(o)

these two occurrences of R^(o) are taken together with the oxygen atomsto which they are bound to form a fused 6-membered oxygen containingring:

It will be appreciated that a variety of other rings can be formed whentwo independent occurrences of R^(o) (or R⁺, or any other variablesimilarly defined herein) are taken together with the atom(s) to whicheach variable is bound and that the examples detailed above are notintended to be limiting.

Unless otherwise stated, structures depicted herein are also meant toinclude all isomeric (e.g., enantiomeric, diastereomeric, and geometric(or conformational)) forms of the structure; for example, the R and Sconfigurations for each asymmetric center, (Z) and (E) double bondisomers, and (Z) and (E) conformational isomers. Therefore, singlestereochemical isomers as well as enantiomeric, diastereomeric, andgeometric (or conformational) mixtures of the present compounds arewithin the scope of the invention. Unless otherwise stated, alltautomeric forms of the compounds of the invention are within the scopeof the invention. Additionally, unless otherwise stated, structuresdepicted herein are also meant to include compounds that differ only inthe presence of one or more isotopically enriched atoms. For example,compounds having the present structures except for the replacement ofhydrogen by deuterium or tritium, or the replacement of a carbon by a¹³C— or ¹⁴C-enriched carbon are within the scope of this invention. Suchcompounds are useful, for example, as analytical tools or probes inbiological assays.

3. Description of Exemplary Compounds:

As described generally above for compounds of formula I, R¹ and R²,taken together with the carbon atoms to which they are bound form anoptionally substituted 5- or 6-membered aryl or heteroaryl ring having0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur.In certain preferred embodiments, R¹ and R², taken together with thecarbon atoms to which they are bound form an optionally substituted 5 or6-membered aryl or heteroaryl ring selected from one of the followinggroups:

wherein W, R^(y), y, and R³ are as defined generally above and inclasses and subclasses herein. It should be understood that theorientation of the rings i to xxi-2 relative to formula I is as follows:

In more preferred embodiments, R¹ and R², taken together with the carbonatoms to which they are bound form an optionally substituted 5- or6-membered aryl or heteroaryl ring selected from phenyl (i), pyridyl(ii-1, ii-2, ii-3, ii-4), pyrazolyl (xiii-1, xiii-2, xiii-3, andxiii-4), oxadiazolyl (xiv), thiophenyl (iv-1, iv-2, and xviii), orthiazolyl (viii-1 and viii-2).

It will be appreciated that the ring formed by R¹ and R² taken togetheris optionally substituted at one or more carbon atoms with y independentoccurrences of —WR^(y), wherein y is 0-5; and is optionally substitutedat one or more substitutable nitrogen atoms with —R³. In preferredembodiments, each occurrence of WR^(y), when present, is independentlyhalogen, CN, NO₂, —N(R′)₂, —CH₂N(R′)₂, —OR′, —CH₂OR′, —SR′, —CH₂SR′,—COOR′, —NRCOR′, —CON(R′)₂, —S(O)₂N(R′)₂, or an optionally substitutedgroup selected from C₁₋₄alkyl, aryl, or heteroaryl. In more preferredembodiments, —WR^(y) groups are each independently F, Cl, Br, I, Me, Et,—CN, —OMe, —SMe, —NMe₂, —NEt₂, —COOMe, —COOH, —OH, —SO₂NH₂, —CON(CH₃)₂,—CO(optionally substituted N-piperazinyl), —CO(N-morpholinyl) (includingoptionally substituted N-morpholinyl), —CO(N-piperidinyl) (includingoptionally substituted N-piperidinyl), —CH₂N(Me)₂, —CH₂N(Et)₂, or anoptionally substituted group selected from C₁₋₄alkoxy, phenyl,phenyloxy, benzyl, benzyloxy, pyridyl, pyrimidinyl, thiophene,N-morpholinyl, N-piperidinyl, N-piperazinyl, or furanyl. In another morepreferred embodiment, —WR^(y) groups are each independently CF₃, F, Cl,Br, I, Me, Et, —CN, —OMe, —SMe, —NMe₂, —NEt₂, —COOMe, —COOH, —OH,—SO₂NH₂, —CON(CH₃)₂, —CO(optionally substituted N-piperazinyl,—CO(N-morpholinyl) (including optionally substituted N-morpholinyl),—CO(N-piperidinyl) (including optionally substituted N-piperidinyl),—CO(pyrrolidinyl) (including optionally substituted pyrrolidinyl),—CO(N(H)-pyrrolidinyl) (including —COIN-pyrrolidinyl), wherein eachpyrrolidinyl is optionally substituted), —CH₂N(Me)₂, —CH₂N(Et)₂, or anoptionally substituted group selected from C₁₋₄alkoxy, phenyl,phenyloxy, benzyl, benzyloxy, pyridyl, pyrimidinyl, thiophene,N-morpholinyl, N-piperidinyl, N-piperazinyl, furanyl, pyrrolidinyl, or—N(H)pyrrolidinyl. In other preferred embodiments, y is 1 and WR^(y) isan optionally substituted aryl or heteroaryl group. Preferredsubstituents for the optionally substituted aryl or heteroaryl groupinclude —VR^(v); wherein V is a bond or is a C₁-C₆ alkylidene chainwherein up to two non-adjacent methylene units of V are independentlyoptionally replaced by CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO,NRCO₂, NRCONR, SO, SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR; and eachoccurrence of R^(v) is independently selected from R′, halogen, NO₂, orCN, or -QR^(x) is ═O, ═S, or ═NR′. In still other embodiments, y is 1and WR^(y) is —CH₂N(R′)₂, —N(R′)₂, or —CON(R′)₂. Most preferred WR^(y)groups and substituents thereof include those shown below in Tables 1-8.In certain embodiments, y is 0.

Preferred T groups, when present, include, but are not limited to,—CH₂—, —CH₂CH₂—, —CO— and —SO₂—. Alternatively, T groups, when presentare —C(H)(CH₃)—, —C(H)(CH₃)CH₂—, —CH₂C(H)(CH₃)—, or —CH₂CH₂CH₂—. Incertain other preferred embodiments, n is 0 and T is absent. In mostpreferred embodiments, T is absent (n=0) or T is —CO—.

As described generally above, Cy¹ is a 3-7 membered saturated, partiallyunsaturated, or fully unsaturated monocyclic ring having 0-3 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or an 8-10membered saturated, partially unsaturated, or fully unsaturated bicyclicring system having 0-5 heteroatoms independently selected from nitrogen,oxygen, or sulfur. In certain embodiments, for compounds of generalformula I, Cy¹ is selected from one of the following groups:

wherein Q, R⁴ and R^(x) are as defined generally above and in classesand subclasses herein, and x is 0-5.

In more preferred embodiments, Cy¹ is selected from phenyl (a), pyridyl(b) (preferably attached in the 2-, 3, or 4-position as shown below byb-i, b-ii, and b-iii), pyrimidinyl (c) (preferably attached in the 2-,4- or 5-position as shown by c-i, c-ii, and c-iii), imidazolyl (g)(preferably attached in the 2-, 4- or 5-position as shown by g-i),thienyl (l), thiazolyl (o) (preferably attached in the 2-position asshown by o-i), cyclohexyl (v), piperazinyl (aa), morpholinyl (bb),thiomorpholinyl (ff), pyrrolidinyl (gg), tetrahydrofuryl (hh),tetrahydrothiofuryl (ii), and cyclopropyl (kk):

In other preferred embodiments, Cy¹ is selected from phenyl (a), pyridyl(b) (preferably attached in the 2-, 3, or 4-position as shown by b-i,b-ii, and b-iii), imidazolyl (g) (preferably attached in the 2-, 4- or5-position as shown by g-i), benzimidazol-2-yl (ll), thiazolyl (o)(preferably attached in the 2-position as shown by o-i),benzthiazol-2-yl (mm), and thienyl (l):

In most preferred embodiments Cy¹ is selected from phenyl (a), andcompounds have the formula I-A:

wherein X, R¹, R², n, T, Q and R^(x) are as defined generally above andin classes and subclasses herein and x is 0-5.

As detailed above, Cy¹ can be optionally substituted with up to 5occurrences of QR^(x). In certain preferred embodiments, x is 0-3, andthus Cy¹ is substituted with 0-3 occurrences of QR^(x). In still otherpreferred embodiments, x is 0 and Cy¹ is unsubstituted.

In preferred embodiments, QR^(x) groups, when present, are eachindependently halogen, CN, NO₂, or an optionally substituted groupselected from C₁₋₄allyl, aryl, aralkyl, —N(R′)₂, —CH₂N(R′)₂, —OR′,—CH₂OR′, —SR′, —CH₂SR′, —COOR′, —NRCOR′, —CON(R′)₂, —SO₂R′, NR′SO₂R′, or—SO₂N(R′)₂. Alternatively, QR^(x) groups, when present, are eachindependently halogen, CN, NO₂, or an optionally substituted groupselected from C₁₋₄alkyl, —CF₃, aryl, heteroaryl, aralkyl, —N(R′)₂,—CH₂N(R′)₂, —OR′, —CH₂OR′, —SR′, —CH₂SR′, —COOR′, —NRCOR′, —CON(R′)₂,—SO₂R′, NR′SO₂R′, SO₂N═R′, or —SO₂N(R′)₂. In more preferred embodiments,QR^(x) groups are each independently Cl, Br, F, CF₃, methyl, ethyl,isopropyl, n-propyl, n-butyl, tert-butyl, NO₂, —OH, —SO₂NH₂, SO₂CH₃,NH₂, SO₂NHCH₃, NHSO₂CH₃, or an optionally substituted group selectedfrom C₁₋₄alkoxy, phenyl, phenyloxy, benzyl, or benzyloxy. Most preferredQR^(x) groups include those shown below in Tables 1-8.

As described generally above, X is an optionally substituted C₁-C₃alkylidene chain wherein one or two non-adjacent methylene units areindependently optionally replaced by CO, CONR′, NR′CO, SO, SO₂, NR′SO₂,SO₂NR′, O, S, or NR′. In certain preferred embodiments, compounds ofspecial interest include those compounds wherein X is an optionallysubstituted methylene group and compounds have the general formula II:

As described generally above, in certain preferred embodiments, Cy¹ isselected from any one of a through mm depicted above (including certainsubsets b-i, c-i, b-ii, b-iii, c-ii, c-iii, g-i, ll, o-i or mm). It willbe appreciated, however, that for compounds of formula II as describedabove, certain additional compounds are of special interest. Forexample, in certain exemplary embodiments, for compounds of generalformula II above, compounds of special interest include those compoundswhere Cy¹ is optionally substituted phenyl, and compounds have theformula II-A:

wherein X, R¹, R², n, x, T, Q and R^(x) are as defined generally aboveand in classes and subclasses herein.

For certain other compounds of interest, R¹ and R², taken together withthe carbon atoms to which they are bound form an optionally substituted5 or 6-membered aryl or heteroaryl ring selected from phenyl (i),pyridyl (ii-1, ii-2, ii-3, ii-4), pyrazolyl (xiii-2), oxadiazolyl (xiv),thiophenyl (iv-1, iv-2, and xviii), or thiazolyl (viii-1 and viii-2),and compounds have one of the structures II-B, II-C, II-D, II-E, II-F,II-G, II-H, II-I, II-J, II-K, II-L, or II-M:

It will be appreciated that certain subclasses of the foregoingcompounds of formulas II and II-A through II-M, are of particularinterest.

For example, in certain preferred embodiments, for compounds of formulasII and II-A through II-M, Cy¹ is phenyl, optionally substituted with 0-3occurrences of QR^(x). In more preferred embodiments for compoundsdescribed above, n is 0, or n is 1 and T is CH₂, —CH₂CH₂—, —CO— or—SO₂—; x is 0-3; and each occurrence of QR^(x) is independently halogen,CN, NO₂, or an optionally substituted group selected from C₁₋₄alkyl,aryl, aralkyl, —N(R′)₂, —CH₂N(R′)₂, —OR′, —CH₂OR′, —SR′, —CH₂SR′,—COOR′, —NRCOR′, —CON(R′)₂, —SO₂R′, NR′SO₂R′, or —SO₂N(R′)₂.Alternatively, T is —C(H)(CH₃)—-C(H)(CH₃)CH₂—, —CH₂C(H)(CH₃)—, or—CH₂CH₂CH₂—.

In more preferred embodiments, QR^(x) groups are each independently Cl,Br, F, CF₃, methyl, ethyl, isopropyl, n-propyl, n-butyl, tert-butyl,NO₂, —OH, —SO₂NH₂, SO₂CH₃, NH₂, SO₂NHCH₃, NHSO₂CH₃, or an optionallysubstituted group selected from C₁₋₄alkoxy, phenyl, phenyloxy, benzyl,or benzyloxy.

In certain preferred embodiments, for compounds of formula II-A, x is0-3; and each occurrence of QR^(x) is independently halogen, CN, NO₂, oran optionally substituted group selected from C₁₋₄alkyl, aryl, aralkyl,—N(R′)₂, —CH₂N(R′)₂, —OR′, —CH₂OR′, —SR′, —CH₂SR′, —COOR′, —NRCOR′,—CON(R′)₂, —SO₂R′, NR′SO₂R′, or —SO₂N(R′)₂. In more preferredembodiments, QR^(x) groups are each independently Cl, Br, F, CF₃,methyl, ethyl, isopropyl, n-propyl, n-butyl, tert-butyl, NO₂, —OH,—SO₂NH₂, SO₂CH₃, NH₂, SO₂NHCH₃, NHSO₂CH₃, or an optionally substitutedgroup selected from C₁₋₄alkoxy, phenyl, phenyloxy, benzyl, or benzyloxy.

In certain preferred embodiments, for compounds of formulae II-B throughII-M, y is 0-3; and each occurrence of WR^(y) is independently halogen,CN, NO₂, or an optionally substituted group selected from C₁₋₄alkyl,aryl, heteroaryl, —N(R′)₂, —CH₂N(R′)₂, —OR′, —CH₂OR′, —SR′, —CH₂SR′,—COOR′, —NRCOR′, —CON(R′)₂, or —S(O)₂N(R)₂. In more preferredembodiments, —WR^(y) groups are each independently F, Cl, Br, I, Me, Et,—CN, —OMe, —SMe, —NMe₂, —NEt₂, —COOMe, —COOH, —OH, —SO₂NH₂, —CON(CH₃)₂,—CO(optionally substituted N-piperazinyl), —CO(N-morpholinyl) (includingoptionally substituted N-morpholinyl), —CO(N-piperidinyl) (includingoptionally substituted N-piperidinyl), —CH₂N(Me)₂, —CH₂N(Et)₂, or anoptionally substituted group selected from C₁₋₄alkoxy, phenyl,phenyloxy, benzyl, benzyloxy, pyridyl, pyrimidinyl, thiophene,N-morpholinyl, N-piperidinyl, N-piperazinyl, or furanyl. Alternatively,—WR^(y) groups are each independently CF₃, F, Cl, Br, I, Me, Et, —CN,—OMe, —SMe, —NMe₂, —NEt₂, —COOMe, —COOH, —OH, —SO₂NH₂, —CON(CH₃)₂,—CO(optionally substituted N-piperazinyl), —CO(N-morpholinyl) (includingoptionally substituted N-morpholinyl), —CO(N-piperidinyl) (includingoptionally substituted N-piperidinyl), —CO(pyrrolidinyl) (includingoptionally substituted pyrrolidinyl), —CO(N(H)pyrrolidinyl) (including—COIN-pyrrolidinyl), wherein each pyrrolidinyl is optionallysubstituted), —CH₂N(Me)₂, —CH₂N(Et)₂, or an optionally substituted groupselected from C₁₋₄alkoxy, phenyl, phenyloxy, benzyl, benzyloxy, pyridyl,pyrimidinyl, thiophene, N-morpholinyl, N-piperidinyl, N-piperazinyl,furanyl, pyrrolidinyl, or N(H)pyrrolidinyl.

In other preferred embodiments, y is 1 and WR^(y) is an optionallysubstituted aryl or heteroaryl group. Preferred substituents for theoptionally substituted aryl or heteroaryl group include —VR^(v); whereinV is a bond or is a C₁-C₆ alkylidene chain wherein up to twonon-adjacent methylene units of V are independently optionally replacedby CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO, NRCO₂, NRCONR, SO,SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR; and each occurrence of R^(v) isindependently selected from R′, halogen, NO₂, or CN, or -QR^(x) is ═O,═S, or ═NR′. In still other embodiments, y is 1 and WR^(y) is—CH₂N(R′)₂, —N(R′)₂, or —CON(R′)₂. Most preferred WR^(y) groups andsubstituents thereof include those shown below in Tables 1-8. In certainembodiments, y is 0.

In still other preferred embodiments, compounds have the general formulaII-H and one occurrence of WR^(y) is an optionally substituted aryl orheteroaryl group, denoted by Ar¹ in formula II-H-i below:

In still other preferred embodiments, Ar¹ is an optionally substitutedphenyl, pyridyl, pyrimidinyl, thiophenyl, or furanyl group. Preferredsubstituents for the optionally substituted aryl or heteroaryl groupinclude —VR^(v); wherein V is a bond or is a C₁-C₆ alkylidene chainwherein up to two non-adjacent methylene units of V are independentlyoptionally replaced by CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO,NRCO₂, NRCONR, SO, SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR; and eachoccurrence of R^(v) is independently selected from R′, halogen, NO₂, orCN, or -QR^(x) is ═O, ═S, or ═NR′.

In certain other embodiments, for compounds of formulae II and II-Athrough II-M, n is 0, or n is 1 and T is CH₂, —CH₂CH₂—, —CO— or —SO₂.Alternatively, T is —C(H)(CH₃)—C(H)(CH₃)CH₂—, —CH₂C(H)(CH₃)—, or—CH₂CH₂CH₂—.

In certain other embodiments, for compounds of formula II and II-Athrough II-M, n is 0, or n is 1 and T is CH₂, —CH₂CH₂—, —CO— or —SO₂—; xis 0-3; and each occurrence of QR^(x) is independently halogen, CN, NO₂,or an optionally substituted group selected from C₁₋₄alkyl, aryl,aralkyl, —N(R′)₂, —CH₂N(R′)₂, —OR′, —CH₂OR′, —SR′, —CH₂SR′, —COOR′,—NRCOR′, —CON(R′)₂, NR′SO₂R′, or —SO₂N(R′)₂. Alternatively, T is—C(H)(CH₃)— —C(H)(CH₃)CH₂—, —CH₂C(H)(CH₃)—, or —CH₂CH₂CH₂—. In morepreferred embodiments, QR^(x) groups are each independently Cl, Br, F,CF₃, methyl, ethyl, isopropyl, n-propyl, n-butyl, tert-butyl, NO₂, —OH,—SO₂NH₂, SO₂CH₃, NH₂, SO₂NHCH₃, NHSO₂CH₃, or an optionally substitutedgroup selected from C₁₋₄alkoxy, phenyl, phenyloxy, benzyl, or benzyloxy.Most preferred QR^(x) groups include those shown below in Tables 1-8.

In other preferred embodiments, X is an optionally substitutedC₂-alkylidene chain wherein zero, one or two methylene units areoptionally replaced by S, O, or —SO₂N(R′)—, and the compounds have oneof the structures III, IV, V, VI, VII, VIII, or IX:

wherein W, R^(y), y, and R′ are as defined generally above and inclasses and subclasses herein; wherein the dashed bond denoted a singleor double bond, as valency permits.

As described generally above, in certain preferred embodiments, Cy¹ isselected from any one of a through mm depicted above (including certainsubsets b-i, c-i, b-ii, b-iii, c-ii, c-iii, g-i, ll, o-i or mm). It willbe appreciated, however, that for compounds of formulae III through VIas described above, certain additional compounds are of specialinterest. For example, in certain exemplary embodiments, for compoundsof general formulae II through VI above, compounds of special interestinclude those compounds where Cy¹ is optionally substituted phenyl, andcompounds have one of formulae III-A through VI-A:

wherein X, R¹, R², n, x, T, Q and R^(x) are as defined generally aboveand in classes and subclasses herein.

In certain preferred embodiments, for compounds of formulas III throughIX, R¹ and R², taken together with the carbon atoms to which they arebound form an optionally substituted 5 or 6-membered aryl or heteroarylring selected from phenyl (i), pyridyl (ii-1, ii-2, ii-3, ii-4),pyrazolyl (xiii-2), oxadiazolyl (xiv) and thiophenyl (iv-1, iv-2, andxviii), or thiazolyl (viii-1 and viii-2). In more preferred embodiments,for compounds of general formula III, R¹ and R², taken together with thecarbon atoms to which they are bound form an optionally substituted 5 or6-membered aryl or heteroaryl ring selected from phenyl (i), pyridyl(ii-1, ii-2, ii-3, ii-4), pyrazolyl (xiii-2), oxadiazolyl (xiv),thiophenyl (iv-1, iv-2, and xviii), or thiazolyl (viii-1 and viii-2),and compounds have one of the formulas III-B, III-C, III-D, III-E,III-F, III-G, III-H, III-I, III-J, III-K, III-L, or III-M:

It should be understood that a substituent on a bond passing throughmore than one ring indicates that any of those rings may be substitutedwith the substituent. That is, a compound of formula III-B may besubstituted on either the cyclohexyl or the phenyl with WR^(y) (whereinthe total number of y1 and y2 is equal to y):

In other preferred embodiments, for compounds of general formula IV, R¹and R², taken together with the carbon atoms to which they are boundform an optionally substituted 5 or 6-membered aryl or heteroaryl ringselected from phenyl (i), pyridyl (ii-1, ii-2, ii-3, ii-4), pyrazolyl(xiii-2), oxadiazolyl (xiv), thiophenyl (iv-1, iv-2, and xviii), orthiazolyl (viii-1 and viii-2), and compounds have one of the formulasIV-B, IV-C, IV-D, IV-E, IV-F, IV-G, IV-H, IV-I, IV-J, IV-K, IV-L, orIV-M:

In other preferred embodiments, for compounds of general formula V, R¹and R², taken together with the carbon atoms to which they are boundform an optionally substituted 5 or 6-membered aryl or heteroaryl ringselected from phenyl (i), pyridyl (ii-1, ii-2, ii-3, ii-4), pyrazolyl(xiii-2), oxadiazolyl (xiv), thiophenyl (iv-1, iv-2, and xviii), orthiazolyl (viii-1 and viii-2), and compounds have one of the formulasV-B, V-C, V-D, V-E, V-F, V-G, V-H, V-I, V-J, V-K, V-L, or V-M:

In other preferred embodiments, for compounds of general formula IX, R¹and R², taken together with the carbon atoms to which they are boundform an optionally substituted 5 or 6-membered aryl or heteroaryl ringselected from phenyl (i), pyridyl (ii-1, ii-2, ii-3, ii-4), pyrazolyl(xiii-2), oxadiazolyl (xiv), thiophenyl (iv-1, iv-2, and xviii), orthiazolyl (viii-1 and viii-2), and compounds have one of the formulasIX-B, IX-C, IX-D, IX-E, IX-F, IX-G, IX-H, IX-I, IX-J, IX-K, IX-L, orIX-M:

It will be appreciated that certain subclasses of the foregoingcompounds are of particular interest.

For example, in certain preferred embodiments, for compounds describeddirectly above Cy¹ is phenyl, optionally substituted with 0-3occurrences of QR^(x). In more preferred embodiments for compoundsdescribed above, n is 0, or n is 1 and T is CH₂, —CH₂CH₂—, —CO— or—SO₂—; x is 0-3; and each occurrence of QR^(x) is independently halogen,CN, NO₂, or an optionally substituted group selected from C₁₋₄alkyl,aryl, aralkyl, —N(R)₂, —CH₂N(R′)₂, —CH₂OR′, —SR′, —CH₂SR′, —COOR′,—NRCOR′, —CON(R′)₂, —SO₂R′, NR′SO₂R′, or —SO₂N(R′)₂. Alternatively, T is—C(H)(CH₃)— —C(H)(CH₃)CH₂—, —CH₂C(H)(CH₃)—, or —CH₂CH₂CH₂—. In morepreferred embodiments, QR^(x) groups are each independently Cl, Br, F,CF₃, methyl, ethyl, isopropyl, n-propyl, n-butyl, tert-butyl, NO₂, —OH,—SO₂NH₂, SO₂CH₃, NH₂, SO₂NHCH₃, NHSO₂CH₃, or an optionally substitutedgroup selected from C₁₋₄alkoxy, phenyl, phenyloxy, benzyl, or benzyloxy.

In certain preferred embodiments, for compounds described directlyabove, y is 0-3; and each occurrence of WR^(y) is independently halogen,CN, NO₂, or an optionally substituted group selected from C₁₋₄alkyl,aryl, heteroaryl, —N(R)₂, —CH₂N(R′)₂, —OR′, —CH₂OR′, —SR′, —CH₂SR′,—COOR′, —NRCOR′, —CON(R)₂, or —S(O)₂N(R′)₂. In more preferredembodiments, —WR^(y) groups are each independently F, Cl, Br, I, Me, Et,—CN, —OMe, —SMe, —NMe₂, —NEt₂, —COOMe, —COOH, —OH, —SO₂NH₂, —CON(CH₃)₂,—CO(optionally substituted N-piperazinyl), —CO(N-morpholinyl) (includingoptionally substituted N-morpholinyl), —CO(N-piperidinyl) (includingoptionally substituted N-piperidinyl), —CH₂N(Me)₂, —CH₂N(Et)₂, or anoptionally substituted group selected from C₁₋₄alkoxy, phenyl,phenyloxy, benzyl, benzyloxy, pyridyl, pyrimidinyl, thiophene,N-morpholinyl, N-piperidinyl, N-piperazinyl, or furanyl. Alternatively,—WR^(y) groups are each independently CF₃, F, Cl, Br, I, Me, Et, —CN,—OMe, —SMe, —NMe₂, —NEt₂, —COOMe, —COOH, —OH, —SO₂NH₂, —CON(CH₃)₂,—CO(optionally substituted N-piperazinyl), —CO(N-morpholinyl) (includingoptionally substituted N-morpholinyl), —CO(N-piperidinyl) (includingoptionally substituted N-piperidinyl), —CO(pyrrolidinyl) (includingoptionally substituted pyrrolidinyl), —CO(N(H)pyrrolidinyl) (including—COIN-pyrrolidinyl), wherein each pyrrolidinyl is optionallysubstituted), —CH₂N(Me)₂, —CH₂N(Et)₂, or an optionally substituted groupselected from C₁₋₄alkoxy, phenyl, phenyloxy, benzyl, benzyloxy, pyridyl,pyrimidinyl, thiophene, N-morpholinyl, N-piperidinyl, N-piperazinyl,furanyl, pyrrolidinyl, or N(H)pyrrolidinyl.

In other preferred embodiments, y is 1 and WR^(y) is an optionallysubstituted aryl or heteroaryl group. Preferred substituents for theoptionally substituted aryl or heteroaryl group include —VR^(v); whereinV is a bond or is a C₁-C₆ alkylidene chain wherein up to twonon-adjacent methylene units of V are independently optionally replacedby CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO, NRCO₂, NRCONR, SO,SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR; and each occurrence of R^(v) isindependently selected from R′, halogen, NO₂, or CN, or -QR^(x) is ═O,═S, or ═NR′. In still other embodiments, y is 1 and WR^(y) is—CH₂N(R′)₂, —N(R′)₂, or —CON(R′)₂. Most preferred WR^(y) groups andsubstituents thereof include those shown below in Tables 1-8. In certainembodiments, y is 0.

In still other preferred embodiments, compounds have the general formulaIII-H, III-L, III-M, IV-H, IV-L, IV-M, V-H, V-L, V-M, IX-H, IX-L, orIX-M, and one occurrence of WR^(y) is an optionally substituted aryl orheteroaryl group, denoted by Ar¹ in one of formulae III-H-i, III-L-i,III-M-i, IV-H-i, IV-L-i, IV-M-i, V-H-i, V-L-i, V-M-i, IX-H-i, IX-L-i, orIX-M-i below:

In still other preferred embodiments, Ar¹ is an optionally substitutedphenyl, pyridyl, pyrimidinyl, thiophenyl, or furanyl group. Preferredsubstituents for the optionally substituted aryl or heteroaryl groupinclude —VR^(v); wherein V is a bond or is a C₁-C₆ alkylidene chainwherein up to two non-adjacent methylene units of V are independentlyoptionally replaced by CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO,NRCO₂, NRCONR, SO, SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR; and eachoccurrence of R^(v) is independently selected from R′, halogen, NO₂, orCN, or -QR^(x) is ═O, ═S, or ═NR′.

In yet other preferred embodiments, compounds have the general formulaIII-H, III-L, III-M, IV-H, IV-L, IV-M, V-H, V-L, V-M, IX-H, IX-L, orIX-M and one occurrence of WR^(y) is —CH₂N(R′)₂ and compounds have oneof the formulae III-H-ii, III-L-ii, III-M-ii, IV-H-ii, IV-L-ii, IV-M-ii,V-H-ii, V-L-ii, V-M-ii, IX-H-ii, IX-L-ii, or IX-M-ii below:

In yet other preferred embodiments, compounds have the general formulaIII-H, III-L, III-M, IV-H, IV-L, IV-M, V-H, V-L, V-M, IX-H, IX-L, orIX-M and one occurrence of WR^(y) is —N(R′)₂ and compounds have one ofthe formulae III-H-iii, III-L-iii, III-M-iii, IV-H-iii, IV-L-iii,IV-M-iii, V-H-iii, V-L-iii, V-M-iii, IX-H-iii, IX-L-iii, or IX-M-iiibelow:

In yet other preferred embodiments, compounds have the general formulaIII-H, III-L, III-M, IV-H, IV-L, IV-M, V-H, V-L, V-M, IX-H, IX-L, orIX-M and one occurrence of WR^(y) is —CON(R′)₂ and compounds have one ofthe formulae III-H-iv, III-L-iv, III-M-iv, IV-H-iv, IV-L-iv, IV-M-iv,V-H-iv, V-L-iv, V-M-iv, IX-H-iv, IX-L-iv, or IX-M-iv below:

In still other preferred embodiments, —CH₂N(R′)₂ is preferably—CH₂N(CH₂)₃, —CH₂N(CH₂CH₃)₂, —CH₂(optionally substituted N-piperazinyl),—CH₂(optionally substituted N-piperidinyl), or —CH₂(optionallysubstituted N-morpholinyl); —N(R′)₂ is preferably —N(CH₂)₃, —N(CH₂CH₃)₂,-optionally substituted N-piperazinyl, optionally substitutedN-piperidinyl, or -optionally substituted N-morpholinyl; and —CON(R′)₂is preferably —CON(CH₂)₃, —CON(CH₂CH₃)₂, —CO(optionally substitutedN-piperazinyl), —CO(optionally substituted N-piperidinyl), or—CO(optionally substituted N-morpholinyl). Alternatively, —CH₂N(R′)₂ ispreferably, —CH₂N(CH₂)₃, —CH₂N(CH₂CH₃)₂, —CH₂(optionally substitutedN-piperazinyl), —CH₂(optionally substituted N-piperidinyl), or—CH₂(optionally substituted N-morpholinyl); —N(R′)₂ is preferably—N(CH₂)₃, —N(CH₂CH₃)₂, -optionally substituted N-piperazinyl, optionallysubstituted N-piperidinyl, or -optionally substituted N-morpholinyl; and—CON(R′)₂ is preferably —CON(CH₂)₃, —CON(CH₂CH₃)₂, —CO(optionallysubstituted N-piperazinyl), —CO(optionally substituted N-piperidinyl),—CO(optionally substituted N-morpholinyl), —CO(optionally substitutedpyrrolidinyl (including optionally substituted N-pyrrolidinyl),—CO(N(H)optionally substituted pyrrolidinyl), optionally substitutedpyrrolidinyl, or —N(H)(optionally substituted pyrrolidinyl).

In certain other embodiments, for compounds described directly above, nis 0, or n is 1 and T is CH₂, —CH₂CH₂—, —CO— or —SO₂. Alternatively, Tis —C(H)(CH₃)— —C(H)(CH₃)CH₂—, —CH₂C(H)(CH₃)—, or —CH₂CH₂CH₂—.

In certain other embodiments, for compounds described directly above nis 0, or n is 1 and T is CH₂, —CH₂CH₂—, —CO— or —SO₂—; x is 0-3; andeach occurrence of QR^(x) is independently halogen, CN, NO₂, or anoptionally substituted group selected from C₁₋₄alkyl, aryl, aralkyl,—N(R′)₂, —CH₂N(R′)₂, —OR′, —CH₂OR′, —SR′, —CH₂SR′, —NRCOR′, —CON(R′)₂,—SO₂R′, NR′SO₂R′, or —SO₂N(R′)₂. Alternatively, T is —C(H)(CH₃)——C(H)(CH₃)CH₂—, —CH₂C(H)(CH₃)—, or —CH₂CH₂CH₂—. In more preferredembodiments, QR^(x) groups are each independently Cl, Br, F, CF₃,methyl, ethyl, isopropyl, n-propyl, n-butyl, tert-butyl, NO₂, —OH,—SO₂NH₂, SO₂CH₃, NH₂, SO₂NHCH₃, NHSO₂CH₃, or an optionally substitutedgroup selected from C₁₋₄alkoxy, phenyl, phenyloxy, benzyl, or benzyloxy.Most preferred QR^(x) groups include those shown below in Tables 1-8.

In certain embodiments, preferred compounds are those of formula IV-H(including the subgenerics thereof). In certain other embodiments,preferred compounds are those of formula IV-M (including the subgenericsthereof).

Another class of special interest includes compounds wherein X is anoptionally substituted C₃ alkylidene moiety and compounds have thegeneral formula X:

As described generally above, in certain preferred embodiments, Cy¹ isselected from any one of a through mm depicted above (including certainsubsets b-i, c-i, b-ii, b-iii, c-ii, c-iii, g-i, ll, o-i or mm). It willbe appreciated, however, that for compounds of formulae III through VIas described above, certain additional compounds are of specialinterest. For example, in certain exemplary embodiments, for compoundsof general formula X above, compounds of special interest include thosecompounds where Cy¹ is optionally substituted phenyl, and compounds haveformula X-A:

In certain preferred embodiments, for compounds of formula X, R¹ and R²,taken together with the carbon atoms to which they are bound form anoptionally substituted 5 or 6-membered aryl or heteroaryl ring selectedfrom phenyl (i), pyridyl (ii-1, ii-2, ii-3, ii-4), pyrazolyl (xiii-2),oxadiazolyl (xiv), thiophenyl (iv-1, iv-2, and xviii), or thiazolyl(viii-1 and viii-2). In more preferred embodiments, for compounds ofgeneral formula VII, R¹ and R², taken together with the carbon atoms towhich they are bound form an optionally substituted 5 or 6-membered arylor heteroaryl ring selected from phenyl (i), pyridyl (ii-1, ii-2, ii-3,ii-4), pyrazolyl (xiii-2), oxadiazolyl (xiv), thiophenyl (iv-1, iv-2,and xviii), or thiazolyl (viii-1 and viii-2), and compounds have one ofthe formulae X-B, X-C, X-D, X-E, X-F, X-G, X-H, X-I, X-J, X-K, X-L, orX-M:

It will be appreciated that certain subclasses of the foregoingcompounds of formulas X-B through X-M are of particular interest.

For example, in certain preferred embodiments, for compounds of formulasX-B through X-M, Cy¹ is phenyl, optionally substituted with 0-3occurrences of QR^(x). In more preferred embodiments for compoundsdescribed above, n is 0, or n is 1 and T is CH₂, —CH₂CH₂—, —CO— or—SO₂—; x is 0-3; and each occurrence of QR^(x) is independently halogen,CN, NO₂, or an optionally substituted group selected from C₁₋₄alkyl,aryl, aralkyl, —N(R′)₂, —CH₂N(R′)₂, —OR′, —CH₂OR′, —SR′, —CH₂SR′,—COOR′, —NRCOR′, —CON(R′)₂, —SO₂R′, NR′SO₂R′, or —SO₂N(R′)₂.Alternatively, T is —C(H)(CH₃)— —C(H)(CH₃)CH₂—, —CH₂C(H)(CH₃)—, or—CH₂CH₂CH₂—. In more preferred embodiments, QR^(x) groups are eachindependently Cl, Br, F, CF₃, methyl, ethyl, isopropyl, n-propyl,n-butyl, tert-butyl, NO₂, —OH, —SO₂NH₂, SO₂CH₃, NH₂, SO₂NHCH₃, NHSO₂CH₃,or an optionally substituted group selected from C₁₋₄alkoxy, phenyl,phenyloxy, benzyl, or benzyloxy.

In certain preferred embodiments, for compounds of formulae X-B throughX-M y is 0-3; and each occurrence of WR^(y) is independently halogen,CN, NO₂, or an optionally substituted group selected from C₁₋₄alkyl,aryl, heteroaryl, —N(R′)₂, —CH₂N(R′)₂, —OR′, —CH₂OR′, —SR′, —CH₂SR′,—COOR′, —NRCOR′, —CON(R′)₂, or —S(O)₂N(R′)₂. In more preferredembodiments, —WR^(y) groups are each independently F, Cl, Br, I, Me, Et,—CN, —OMe, —SMe, —NMe₂, —NEt₂, —COOMe, —COOH, —OH, —SO₂NH₂, —CON(CH₃)₂,—CO(optionally substituted N-piperazinyl), —CO(N-morpholinyl),—CO(N-piperidinyl), —CH₂N(Me)₂, —CH₂N(Et)₂, or an optionally substitutedgroup selected from C₁₋₄alkoxy, phenyl, phenyloxy, benzyl, benzyloxy,pyridyl, pyrimidinyl, thiophene, N-morpholinyl, N-piperidinyl,N-piperazinyl, or furanyl. Alternatively, —WR^(Y) groups are eachindependently CF₃, F, Cl, Br, I, Me, Et, —CN, —OMe, —SMe, —NMe₂, —NEt₂,—COOMe, —COOH, —OH, —SO₂NH₂, —CON(CH₃)₂, —CO(optionally substitutedN-piperazinyl), —CO(N-morpholinyl) (including optionally substitutedmorpholinyl), —CO(N-piperidinyl) (including optionally substitutedpiperidinyl), —CO(pyrrolidinyl) (including optionally substitutedpyrrolidinyl), —CO(N(H)pyrrolidinyl) (including —COIN-pyrrolidinyl),wherein each pyrrolidinyl is optionally substituted), —CH₂N(Me)₂,—CH₂N(Et)₂, or an optionally substituted group selected from C₁₋₄alkoxy,phenyl, phenyloxy, benzyl, benzyloxy, pyridyl, pyrimidinyl, thiophene,N-morpholinyl, N-piperidinyl, N-piperazinyl, furanyl, pyrrolidinyl, orN(H)pyrrolidinyl.

In other preferred embodiments, y is 1 and WR^(y) is an optionallysubstituted aryl or heteroaryl group. Preferred substituents for theoptionally substituted aryl or heteroaryl group include —VR^(v); whereinV is a bond or is a C₁-C₆ alkylidene chain wherein up to twonon-adjacent methylene units of V are independently optionally replacedby CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO, NRCO₂, NRCONR, SO,SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR; and each occurrence of R^(v) isindependently selected from R′, halogen, NO₂, or CN, or -QR^(x) is ═O,═S, or ═NR′. In still other embodiments, y is 1 and WR^(y) is—CH₂N(R′)₂, —N(R′)₂, or —CON(R′)₂. Most preferred WR^(y) groups andsubstituents thereof include those shown below in Tables 1-8. In certainembodiments, y is 0.

In still other preferred embodiments, compounds have one of the generalformulae X-H or X-L and one occurrence of WR^(y) is an optionallysubstituted aryl or heteroaryl group, denoted by Ar¹ in formula X-H-iand X-L-i below:

In still other preferred embodiments, Ar¹ is an optionally substitutedphenyl, pyridyl, pyrimidinyl, thiophenyl, or furanyl group. Preferredsubstituents for the optionally substituted aryl or heteroaryl groupinclude —VR^(v); wherein V is a bond or is a C₁-C₆ alkylidene chainwherein up to two non-adjacent methylene units of V are independentlyoptionally replaced by CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO,NRCO₂, NRCONR, SO, SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR; and eachoccurrence of R^(v) is independently selected from R′, halogen, NO₂, orCN, or -QR^(x) is ═O, ═S, or ═NR′. In still other embodiments, y is 1and WR^(y) is —CH₂N(R′)₂, —N(R′)₂, or —CON(R′)₂.

In still other preferred embodiments, compounds have one of the generalformulae X-H or X-L and W is —CH₂—, a bond, or —CO—, and R^(y) is—N(R′)₂ and compounds have one of the formulae X-H-ii and X-L-ii below:

In certain other embodiments, for compounds described directly above, nis 0, or n is 1 and T is CH₂, —CH₂CH₂—, —CO— or —SO₂. Alternatively, Tis —C(H)(CH₃)— —C(H)(CH₃)CH₂—, —CH₂C(H)(CH₃)—, or —CH₂CH₂CH₂—.

In certain other embodiments, for compounds described directly above, nis 0, or n is 1 and T is CH₂, —CH₂CH₂—, —CO— or —SO₂—; x is 0-3; andeach occurrence of QR^(x) is independently halogen, CN, NO₂, or anoptionally substituted group selected from C₁₋₄alkyl, aryl, aralkyl,—N(R′)₂, —CH₂N(R′)₂, —OR′, —CH₂OR′, —SR′, —CH₂SR′, —COOR′, —NRCOR′,—CON(R′)₂, —SO₂R′, NR′SO₂R′, or —SO₂N(R′)₂. Alternatively, T is—C(H)(CH₃)— —C(H)(CH₃)CH₂—, —CH₂C(H)(CH₃)—, or —CH₂CH₂CH₂—. In morepreferred embodiments, QR^(x) groups are each independently Cl, Br, F,CF₃, methyl, ethyl, isopropyl, n-propyl, n-butyl, tert-butyl, NO₂, —OH,—SO₂NH₂, SO₂CH₃, NH₂, SO₂NHCH₃, NHSO₂CH₃, or an optionally substitutedgroup selected from C₁₋₄alkoxy, phenyl, phenyloxy, benzyl, or benzyloxy.Most preferred QR^(x) groups include those shown below in Tables 1-8.

4. General Synthetic Methodology:

The compounds of this invention may be prepared in general by methodsknown to those skilled in the art for analogous compounds, asillustrated by the general scheme below, and the preparative examplesthat follow. In these schemes, the variables are as defined in theschemes or may be readily derived from the compounds of this invention.

Scheme I above shows a general synthetic route that is used forpreparing the compounds 7 of this invention when Cy¹ and WR^(y) are asdescribed herein. The bromothiophene 2 may be prepared by methodssubstantially similar to those described by Pinna, et al, Eur. J. Med.Chem. Chim. Ther. 1994, 29, 447. Intermediate 3 is prepared according toScheme I step (b). Compound 3 is treated with N-substituted guanidine 4according to step (c). Intermediate guanidines of formula 4 may beprepared by reaction of the corresponding amine Cy¹NH₂ with cyanamide bymethods substantially similar to those described by Kaempf, et al, Chem.Ber. 1904, 32, 1682. This reaction is amenable to a variety ofN-substituted guanidines to form compounds of formula 5.

The formation of the biaryl link derivatives 7 is achieved by treatingthe bromide 5 with a boronic acid derivatives in the presence ofpalladium as a catalyst by using the Suzuki coupling methods that arewell known in the art. The reaction is amenable to a variety ofsubstituted aryl or heteroaryl boronic acids.

Scheme II above shows another general synthetic route that has been usedfor preparing the compounds 7 of this invention when Cy¹ and WR^(y) areas described herein. The formation of the biaryl link derivatives 8 isachieved by treating the bromide 2 with a boronic acid derivative in thepresence of palladium as a catalyst by using the Suzuki coupling methodsthat are well known in the art. The reaction is amenable to a variety ofsubstituted aryl or heteroaryl boronic acids. Intermediate 9 is preparedaccording to Scheme II step (b). Compound 9 is treated withN-substituted guanidine 4 according to step (c). Intermediate guanidinesof formula 4 may be prepared by reaction of the corresponding amineCy¹NH₂ with cyanamide by methods substantially similar to thosedescribed by Kaempf, et al, Chem. Ber. 1904, 32, 1682. This reaction isamenable to a variety of N-substituted guanidines to form compounds offormula 7.

Scheme III above shows a general synthetic route that is used forpreparing compounds of formula 14 of this invention where Cy¹ is asdescribed herein. Intermediate 10 is prepared fromthiophene-3-carboxylate 8 in a two steps sequence via the Weinreb amide9. Compound 10 is then treated, in alkali medium, withcarboxymethoxylamine hemihydrochloride to form intermediate 11. Thecyclisation of 11 is achieved using the method described by Forrester,et al, J. Chem. Soc., Perkin Trans. 1 1981, 984. Intermediate 13 issynthesized according to Scheme III step (e). Compound 13 is treatedwith N-substituted guanidine 4 according to step (f). This reaction isamenable to a variety of N-substituted guanidines to form compounds offormula 14.

Table 1 below depicts exemplary compounds prepared according to thegeneral methods described in Schemes I, II, and III.

TABLE 1 Examples of Compounds of Formula I:

I-1

I-2

I-3

I-4

I-5

I-6

I-7

I-8

I-9

I-10

I-11

I-12

I-13

I-14

I-15

I-16

I-17

I-18

I-19

I-20

I-21

I-22

I-23

I-24

I-25

I-26

I-27

I-28

I-29

I-30

I-31

I-32

I-33

I-34

I-35

I-36

I-37

I-38

I-39

I-40

I-41

I-42

I-43

I-44

I-45

I-46

I-47

I-48

I-49

I-50

Scheme IV above shows a general synthetic route that has been used forpreparing compounds 16 of this invention when Cy¹ and WR^(y) are asdescribed herein. The oxidation of 16 is achieved by treating thetricycle 15 with DDQ according to step (a) of Scheme IV.

Table 2 below depicts exemplary compounds prepared according to thegeneral method described in Scheme IV.

TABLE 2 Examples of Compounds of Formula II:

II-1

II-2

II-3

II-4

II-5

II-6

II-7

II-8

II-9

II-10

II-11

II-12

II-13

II-14

II-15

II-16

II-17

II-18

II-19

II-20

II-21

II-22

II-23

II-24

II-25

II-26

II-27

II-28

II-29

II-30

II-31

II-32

II-33

II-34

II-35

II-36

II-37

II-38

II-39

Scheme V above shows a general synthetic route that is used forpreparing the compounds 22 of this invention when Cy¹ and WR^(y) is asdescribed herein. The 2-substituted-5,6-dihydro-4H-benzothiazol-7-ones20 may be prepared by methods described in the literature by Lehmann, etal, Z. Chem 1967, 7, 422. Intermediate 21 is prepared according toScheme V step (a). Compound 21 is treated with N-substituted guanidine 4according to step (b). This reaction is amenable to a variety ofN-substituted guanidines to form compounds of formula 22.

Table 3 below depicts exemplary compounds prepared according to thegeneral method described in Scheme V.

TABLE 3 Examples of Compounds of Formula III: III-1

III-2

III-3

III-4

III-5

III-6

III-7

III-8

III-9

III-10

III-11

III-12

III-13

III-14

III-15

III-16

III-17

III-18

III-19

III-20

III-21

III-22

III-23

III-24

III-25

III-26

III-27

III-28

III-29

III-30

III-31

III-32

III-33

III-34

III-35

III-36

III-37

III-38

III-39

III-40

Scheme VI above shows a general synthetic route that is used forpreparing the compounds 25 of this invention when Cy¹ and WR^(y) are asdescribed herein. Compounds of formula 23 may be prepared by methodssubstantially similar to those described in the literature by Maillard,et al. Eur. J. Med. Chem. Chim. Ther. 1984, 19, 451. Intermediate 24 isprepared according to Scheme VI step (a). Compound 24 is treated withN-substituted guanidine 4 according to step (b). This reaction isamenable to a variety of N-substituted guanidines to form compounds offormula 25.

Scheme VII above shows another general synthetic route that is used forpreparing the compounds 25 of this invention when Cy¹ and WR^(y) are asdescribed herein. The 2-(tert-butyldimethylsilanyloxy)-cyclohex-2-enone26 is well known in the literature (Crimmins, et al. Tetrahedron 1997,53, 8963). Intermediate 27 is prepared according to Scheme VII step (a).Compound 27 is treated with N-substituted guanidine 4 according to step(b). This reaction is amenable to a variety of N-substituted guanidinesto form compounds of formula 28. After a deprotection of 28 to afford29, this latest is treated with bromine to form compounds of formula 30.Intermediate 30 is treated with thioamide 19 according to Scheme VIIstep (e). This reaction is amenable to a variety of thioamides to formcompounds of structure 25.

Table 4 below depicts exemplary compounds prepared according to thegeneral method described in Schemes VI and VII.

TABLE 4 Examples of Compounds of Formula IV: IV-1

IV-2

IV-3

IV-4

IV-5

IV-6

IV-7

IV-8

IV-9

IV-10

IV-11

IV-12

IV-13

IV-14

IV-15

IV-16

IV-17

IV-18

IV-19

IV-20

IV-21

IV-22

IV-23

IV-24

IV-25

IV-26

IV-27

IV-28

IV-29

IV-30

IV-31

IV-32

IV-33

Scheme VIII above shows a general synthetic route that is used forpreparing compounds 33 of this invention when Cy¹ is as describedherein. The 7H-thieno[2,3-c]thiopyran-4-one 31 (WR^(y)═H) may beprepared by methods described by Mandal, et al, J. Chem. Soc., PerkinTrans. 1 1999, 2639. Intermediate 32 is synthesized according the schemeVIII step (a). Compound 32 is treated with N-substituted guanidine 4according to step (b). This reaction is amenable to a variety ofN-substituted guanidines to form compounds of formula 33.

Scheme IX above shows a general synthetic approach that is used forpreparing compounds 38 and 39 of this invention. The formation ofcompounds of structure 34 was achieved by methods substantially similarto those described by Lo, et al, J. Am. Chem. Soc. 1954, 76, 4166 and byBehringer, et al, Chem. Ber. 1966, 3309. Intermediate 35 is preparedaccording to Scheme IX step (a). The tert-butyl ester 36, prepared bymethods well known in the art was treated with N,N-dimethylformamidedimethylacetal complex to form intermediate 37. Compound 37 is treatedwith N-substituted guanidine 4 according to step (d). This reaction isamenable to a variety of N-substituted guanidines to form compounds offormula 38 of Scheme IX. The aromatization of compound 38 where G is acarbon is achieved in presence of DDQ.

Scheme X above shows a general method for preparing compounds of formula42 of this invention when X and when Cy¹ are as described herein. Eachof the above steps is well known to one of skill in the art. OK

Scheme XI above shows a general synthetic approach that is used forpreparing compounds 75 of this invention. Intermediate 70, prepared bycyclisation of compound of formula 69 in presence ofS-methylisothiouronium sulphate, was aromatized with DDQ according toScheme XI step (b). The acid 73, obtained by subsequent oxidation to thesulfone 72 and saponification, is treated with an amine RR′NH in acoupling reaction step well known to one of skill in the art. Thisreaction is amenable to a variety of amine RR′NH to form compounds offormula 74 of Scheme XI. The displacement of the sulfone by an amineCy¹NH₂ is achieved according to Scheme XI step (f). This reaction isamenable to a variety of amine Cy¹NH₂ to form compounds of formula 75 ofScheme XI.

Table 5 below depicts exemplary compounds prepared according to thegeneral method described in Schemes VIII, IX, X and XI.

TABLE 5 Examples of Compounds of Formula V: V-1

V-2

V-3

V-4

V-5

V-6

V-7

V-8

V-9

V-10

V-11

V-12

V-13

V-14

V-15

V-16

V-17

V-18

V-19

V-20

V-21

V-22

V-23

V-24

V-25

V-26

V-27

V-28

V-29

V-30

V-31

V-32

V-33

V-34

V-35

V-36

V-37

V-38

V-39

V-40

V-41

V-42

V-43

V-44

V-45

V-46

V-47

V-48

V-49

V-50

V-51

V-52

V-53

V-54

V-55

V-56

V-57

V-58

V-59

V-60

V-61

V-62

V-63

V-64

V-65

V-66

V-67

V-68

V-69

V-70

V-71

V-72

V-73

V-74

V-75

V-76

V-77

V-78

V-79

V-80

V-81

V-82

V-83

V-84

V-85

V-86

V-87

V-88

V-89

V-90

V-91

V-92

V-93

V-94

V-95

V-96

V-97

V-98

V-99

V-100

V-101

V-102

V-103

V-104

V-105

V-106

V-107

V-108

V-109

V-110

V-111

V-112

V-113

V-114

V-115

V-116

V-117

V-118

V-119

V-120

V-121

V-122

V-123

V-124

V-125

V-126

V-127

V-128

V-129

V-130

V-131

V-132

V-133

V-134

V-135

V-136

V-137

V-138

V-139

V-140

V-141

V-142

V-143

V-144

V-145

V-146

V-147

V-148

V-149

V-150

V-151

V-152

V-153

V-154

V-155

V-156

V-157

V-158

V-159

V-160

V-161

V-162

V-163

V-164

V-165

V-166

V-167

V-168

V-169

V-170

V-171

V-172

V-173

V-174

V-175

V-176

V-177

V-178

V-179

V-180

V-181

V-182

Scheme XII above shows a general synthetic route that is used forpreparing compounds of formula 45 of this invention where X and Cy¹ areas described above. Each of the above steps is well known to one ofskill in the art.

Table 6 below depicts exemplary compounds prepared according to thegeneral method described in Scheme XII.

TABLE 6 Examples of Compounds of Formula VI: VI-1

VI-2

VI-3

VI-4

VI-5

VI-6

VI-7

VI-8

VI-9

VI-10

VI-11

VI-12

VI-13

VI-14

VI-15

VI-16

VI-17

VI-18

VI-19

VI-20

VI-21

VI-22

VI-23

VI-24

VI-25

VI-26

VI-27

Scheme XIII above shows a general synthetic route that is used forpreparing compounds of formula 58 of this invention where X is definedgenerally herein and where Cy¹, WR^(Y), and y are as described herein.Each of the above steps is well known to one of skill in the art.

Table 7 below depicts exemplary compounds prepared according to thegeneral method described in Scheme XIII.

TABLE 7 Examples of Compounds of Formula VII: VII-1

VII-2

VII-3

VII-4

VII-5

VII-6

VII-7

VII-8

VII-9

VII-10

VII-11

VII-12

VII-13

VII-14

VII-15

VII-16

VII-17

VII-18

VII-19

VII-20

VII-21

VII-22

VII-23

VII-24

VII-25

VII-26

VII-27

VII-28

VII-29

VII-30

VII-31

VII-32

VII-33

VII-34

VII-35

VII-36

VII-37

Scheme XIV above shows a general synthetic route that is used forpreparing compounds of formula 62 of this invention where Cy¹ and R³ areas described herein. Each of the above steps is well known to one ofskill in the art.

Scheme XV above shows a general synthetic route that is used forpreparing compounds of formula 65 of this invention where Cy¹ is asdescribed herein. Each of the above steps is well known to one of skillin the art.

Scheme XVI above shows a general synthetic route that is used forpreparing compounds of formula 68 of this invention where Cy¹, WR^(Y),and y are as described herein. Each of the above steps is well known toone of skill in the art.

Table 8 below depicts exemplary compounds prepared according to thegeneral method depicted in Schemes XIV, XV, and XVI.

TABLE 8 Examples of Compounds of Formula VIII: VIII-1

VIII-2

VIII-3

VIII-4

VIII-5

VIII-6

VIII-7

VIII-8

VIII-9

Although certain exemplary embodiments are depicted and described aboveand herein, it will be appreciated that compounds of the invention canbe prepared according to the methods described generally above usingappropriate starting materials by methods generally available to one ofordinary skill in the art. Accordingly, another embodiment of thisinvention provides a process for preparing a compound of this inventionaccording to these methods.

5. Uses, Formulation and Administration

Pharmaceutically Acceptable Compositions

As discussed above, the present invention provides compounds that areinhibitors of protein kinases, and thus the present compounds are usefulfor the treatment of diseases, disorders, and conditions including, butnot limited to acute-myelogenous leukemia (AML), acute lymphocyticleukemia (ALL), mastocytosis, gastrointestinal stromal tumor (GIST),cancer including but not limited to colon, breast, hepatic andpancreatic cancer, certain B-cell leukemias and lymphomas,neurodegenerative disorders, diabetes, CNS disorders such as manicdepressive disorder and neurodegenerative diseases, cardiomyocytehypertrophy, immune system dysfunction, bone remodeling diseases,allergic disorders such as asthma, autoimmune diseases such astransplant rejection, rheumatoid arthritis, amyotrophic lateralsclerosis and multiple sclerosis. Accordingly, in another aspect of thepresent invention, pharmaceutically acceptable compositions areprovided, wherein these compositions comprise any of the compounds asdescribed herein, and optionally comprise a pharmaceutically acceptablecarrier, adjuvant or vehicle. In certain embodiments, these compositionsoptionally further comprise one or more additional therapeutic agents.

It will also be appreciated that certain of the compounds of presentinvention can exist in free form for treatment, or where appropriate, asa pharmaceutically acceptable derivative thereof. According to thepresent invention, a pharmaceutically acceptable derivative includes,but is not limited to, pharmaceutically acceptable salts, esters, saltsof such esters, or any other adduct or derivative which uponadministration to a patient in need is capable of providing, directly orindirectly, a compound as otherwise described herein, or a metabolite orresidue thereof.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgement,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. A“pharmaceutically acceptable salt” means any non-toxic salt or salt ofan ester of a compound of this invention that, upon administration to arecipient, is capable of providing, either directly or indirectly, acompound of this invention or an inhibitorily active metabolite orresidue thereof. As used herein, the term “inhibitorily activemetabolite or residue thereof” means that a metabolite or residuethereof is also an inhibitor of a GSK-3, SYK, Aurora-2, CDK-2, JAK-3,LCK, SRC, or Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk)protein kinases kinase.

Pharmaceutically acceptable salts are well known in the art. Forexample, S. M. Berge et al., describe pharmaceutically acceptable saltsin detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporatedherein by reference. Pharmaceutically acceptable salts of the compoundsof this invention include those derived from suitable inorganic andorganic acids and bases. Examples of pharmaceutically acceptable,nontoxic acid addition salts are salts of an amino group formed withinorganic acids such as hydrochloric acid, hydrobromic acid, phosphoricacid, sulfuric acid and perchloric acid or with organic acids such asacetic acid, oxalic acid, maleic acid, tartaric acid, citric acid,succinic acid or malonic acid or by using other methods used in the artsuch as ion exchange. Other pharmaceutically acceptable salts includeadipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate,bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Salts derived from appropriate bases include alkali metal,alkaline earth metal, ammonium and N⁺(C₁₋₄alkyl)₄ salts. This inventionalso envisions the quaternization of any basic nitrogen-containinggroups of the compounds disclosed herein. Water or oil-soluble ordispersable products may be obtained by such quaternization.Representative alkali or alkaline earth metal salts include sodium,lithium, potassium, calcium, magnesium, and the like. Furtherpharmaceutically acceptable salts include, when appropriate, nontoxicammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, loweralkyl sulfonate and aryl sulfonate.

As described above, the pharmaceutically acceptable compositions of thepresent invention additionally comprise a pharmaceutically acceptablecarrier, adjuvant, or vehicle, which, as used herein, includes any andall solvents, diluents, or other liquid vehicle, dispersion orsuspension aids, surface active agents, isotonic agents, thickening oremulsifying agents, preservatives, solid binders, lubricants and thelike, as suited to the particular dosage form desired. Remington'sPharmaceutical Sciences, Sixteenth Edition, E. W. Martin (MackPublishing Co., Easton, Pa., 1980) discloses various carriers used informulating pharmaceutically acceptable compositions and knowntechniques for the preparation thereof. Except insofar as anyconventional carrier medium is incompatible with the compounds of theinvention, such as by producing any undesirable biological effect orotherwise interacting in a deleterious manner with any othercomponent(s) of the pharmaceutically acceptable composition, its use iscontemplated to be within the scope of this invention. Some examples ofmaterials which can serve as pharmaceutically acceptable carriersinclude, but are not limited to, ion exchangers, alumina, aluminumstearate, lecithin, serum proteins, such as human serum albumin, buffersubstances such as phosphates, glycine, sorbic acid, or potassiumsorbate, partial glyceride mixtures of saturated vegetable fatty acids,water, salts or electrolytes, such as protamine sulfate, disodiumhydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zincsalts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone,polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, woolfat, sugars such as lactose, glucose and sucrose; starches such as cornstarch and potato starch; cellulose and its derivatives such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate; powderedtragacanth; malt; gelatin; talc; excipients such as cocoa butter andsuppository waxes; oils such as peanut oil, cottonseed oil; saffloweroil; sesame oil; olive oil; corn oil and soybean oil; glycols; such apropylene glycol or polyethylene glycol; esters such as ethyl oleate andethyl laurate; agar; buffering agents such as magnesium hydroxide andaluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline;Ringer's solution; ethyl alcohol, and phosphate buffer solutions, aswell as other non-toxic compatible lubricants such as sodium laurylsulfate and magnesium stearate, as well as coloring agents, releasingagents, coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator.

Uses of Compounds and Pharmaceutically Acceptable Compositions

In yet another aspect, a method for the treatment or lessening theseverity of a GSK-3, SYK, Aurora-2, CDK-2, JAK-3, LCK, SRC, and/or Tecfamily (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk)-mediated diseases isprovided comprising administering an effective amount of a compound, ora pharmaceutically acceptable composition comprising a compound to asubject in need thereof. In certain embodiments of the present inventionan “effective amount” of the compound or pharmaceutically acceptablecomposition is that amount effective for a GSK-3, SYK, Aurora-2, CDK-2,JAK-3, LCK, SRC, and/or Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx,Txk/Rlk)-mediated disease. The compounds and compositions, according tothe method of the present invention, may be administered using anyamount and any route of administration effective for treating orlessening the severity of a GSK-3, SYK, Aurora-2, CDK-2, JAK-3, LCK,SRC, and/or Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx,Txk/Rlk)-mediated disease. The exact amount required will vary fromsubject to subject, depending on the species, age, and general conditionof the subject, the severity of the infection, the particular agent, itsmode of administration, and the like. The compounds of the invention arepreferably formulated in dosage unit form for ease of administration anduniformity of dosage. The expression “dosage unit form” as used hereinrefers to a physically discrete unit of agent appropriate for thepatient to be treated. It will be understood, however, that the totaldaily usage of the compounds and compositions of the present inventionwill be decided by the attending physician within the scope of soundmedical judgment. The specific effective dose level for any particularpatient or organism will depend upon a variety of factors including thedisorder being treated and the severity of the disorder; the activity ofthe specific compound employed; the specific composition employed; theage, body weight, general health, sex and diet of the patient; the timeof administration, route of administration, and rate of excretion of thespecific compound employed; the duration of the treatment; drugs used incombination or coincidental with the specific compound employed, andlike factors well known in the medical arts. The term “patient”, as usedherein, means an animal, preferably a mammal, and most preferably ahuman.

The pharmaceutically acceptable compositions of this invention can beadministered to humans and other animals orally, rectally, parenterally,intracistemally, intravaginally, intraperitoneally, topically (as bypowders, ointments, or drops), bucally, as an oral or nasal spray, orthe like, depending on the severity of the infection being treated. Incertain embodiments, the compounds of the invention may be administeredorally or parenterally at dosage levels of about 0.01 mg/kg to about 50mg/kg and preferably from about 1 mg/kg to about 25 mg/kg, of subjectbody weight per day, one or more times a day, to obtain the desiredtherapeutic effect.

Liquid dosage forms for oral administration include, but are not limitedto, pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof. Besides inert diluents,the oral compositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a compound of the present invention,it is often desirable to slow the absorption of the compound fromsubcutaneous or intramuscular injection. This may be accomplished by theuse of a liquid suspension of crystalline or amorphous material withpoor water solubility. The rate of absorption of the compound thendepends upon its rate of dissolution that, in turn, may depend uponcrystal size and crystalline form. Alternatively, delayed absorption ofa parenterally administered compound form is accomplished by dissolvingor suspending the compound in an oil vehicle. Injectable depot forms aremade by forming microencapsule matrices of the compound in biodegradablepolymers such as polylactide-polyglycolide. Depending upon the ratio ofcompound to polymer and the nature of the particular polymer employed,the rate of compound release can be controlled. Examples of otherbiodegradable polymers include poly(orthoesters) and poly(anhydrides).Depot injectable formulations are also prepared by entrapping thecompound in liposomes or microemulsions that are compatible with bodytissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like. The solid dosage forms of tablets, dragees, capsules, pills,and granules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes. Solid compositions of a similartype may also be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polyethylene glycols and the like.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositions thatcan be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, and eye drops are also contemplatedas being within the scope of this invention. Additionally, the presentinvention contemplates the use of transdermal patches, which have theadded advantage of providing controlled delivery of a compound to thebody. Such dosage forms can be made by dissolving or dispensing thecompound in the proper medium. Absorption enhancers can also be used toincrease the flux of the compound across the skin. The rate can becontrolled by either providing a rate controlling membrane or bydispersing the compound in a polymer matrix or gel.

As described generally above, the compounds of the invention are usefulas inhibitors of protein kinases. In one embodiment, the compounds andcompositions of the invention are inhibitors of one or more of GSK-3,SYK, Aurora-2, CDK-2, JAK-3, LCK, SRC, or Tec family (e.g., Tec, Btk,Itk/Emt/Tsk, Bmx, Txk/Rlk) kinase, and thus, without wishing to be boundby any particular theory, the compounds and compositions areparticularly useful for treating or lessening the severity of a disease,condition, or disorder where activation of one or more of GSK-3, SYK,Aurora-2, CDK-2, JAK-3, LCK, SRC, and/or Tec family (e.g., Tec, Btk,Itk/Emt/Tsk, Bmx, Txk/Rlk) kinase is implicated in the disease,condition, or disorder. When activation of GSK-3, SYK, Aurora-2, CDK-2,JAK-3, LCK, SRC, and/or Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx,Txk/Rlk) is implicated in a particular disease, condition, or disorder,the disease, condition, or disorder may also be referred to as a “GSK-3,SYK, Aurora-2, CDK-2, JAK-3, LCK, SRC, and/or Tec family (e.g., Tec,Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk)-mediated disease” or disease symptom.Accordingly, in another aspect, the present invention provides a methodfor treating or lessening the severity of a disease, condition, ordisorder where activation or one or more of GSK-3, SYK, Aurora-2, CDK-2,JAK-3, LCK, SRC, and/or Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx,Txk/Rlk) is implicated in the disease state.

The activity of a compound utilized in this invention as an inhibitor ofGSK-3, SYK, Aurora-2, CDK-2, JAK-3, LCK, SRC, and/or Tec family (e.g.,Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk), may be assayed in vitro, in vivoor in a cell line. In vitro assays include assays that determineinhibition of either the phosphorylation activity or ATPase activity ofactivated GSK-3, SYK, Aurora-2, CDK-2, JAK-3, LCK, SRC, and/or Tecfamily (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk). Alternate in vitroassays quantitate the ability of the inhibitor to bind to GSK-3, SYK,Aurora-2, CDK-2, JAK-3, LCK, SRC, and/or Tec family (e.g., Tec, Btk,Itk/Emt/Tsk, Bmx, Txk/Rlk). Inhibitor binding may be measured byradiolabelling the inhibitor prior to binding, isolating theinhibitor/GSK-3, SYK, Aurora-2, CDK-2, JAK-3, LCK, SRC, and/or Tecfamily (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk), complex anddetermining the amount of radiolabel bound. Alternatively, inhibitorbinding may be determined by running a competition experiment where newinhibitors are incubated with GSK-3, SYK, Aurora-2, CDK-2, JAK-3, LCK,SRC, and/or Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) boundto known radioligands.

The term “measurably inhibit”, as used herein means a measurable changein GSK-3, SYK, Aurora-2, CDK-2, JAK-3, LCK, SRC, and/or Tec family(e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) activity between a samplecomprising said composition and a GSK-3, SYK, Aurora-2, CDK-2, JAK-3,LCK, SRC, and/or Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk)kinase and an equivalent sample comprising GSK-3, SYK, Aurora-2, CDK-2,JAK-3, LCK, SRC, and/or Tec family (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx,Txk/Rlk) kinase in the absence of said composition.

The term “Aurora-2-mediated disease” or “Aurora-2-mediated condition”,as used herein, means any disease or other deleterious condition inwhich Aurora is known to play a role. The terms “Aurora-2-mediateddisease” or “Aurora-2-mediated condition” also mean those diseases orconditions that are alleviated by treatment with an Aurora-2 inhibitor.Such conditions include, without limitation, colon, breast, stomach, andovarian cancer. The term “Aurora-2-mediated disease”, as used herein,means any disease or other deleterious condition or disease in whichAurora-2 is known to play a role. Such diseases or conditions include,without limitation, cancers such as colon and breast cancer.

The terms “CDK-2-mediated disease” or “CDK-2-mediated condition”, asused herein, mean any disease or other deleterious condition in whichCDK-2 is known to play a role. The terms “CDK-2-mediated disease” or“CDK-2-mediated condition” also mean those diseases or conditions thatare alleviated by treatment with a CDK-2 inhibitor. Such conditionsinclude, without limitation, cancer, Alzheimer's disease, restenosis,angiogenesis, glomerulonephritis, cytomegalovirus, HIV, herpes,psoriasis, atherosclerosis, alopecia, and autoimmune diseases such asrheumatoid arthritis. See Fischer, P. M. and Lane, D. P. CurrentMedicinal Chemistry, 2000, 7, 1213-1245; Mani, S., Wang, C., Wu, K.,Francis, R. and Pestell, R. Exp. Opin. Invest. Drugs 2000, 9, 1849; Fry,D. W. and Garrett, M. D. Current Opinion in Oncologic, Endocrine &Metabolic Investigational Drugs 2000, 2, 40-59.

The term “GSK-3-mediated disease” as used herein, means any disease orother deleterious condition or disease in which GSK-3 is known to play arole. Such diseases or conditions include, without limitation,autoimmune diseases, inflammatory diseases, metabolic, neurological andneurodegenerative diseases, cardiovascular diseases, allergy, asthma,diabetes, Alzheimer's disease, Huntington's disease, Parkinson'sdisease, AIDS-associated dementia, amyotrophic lateral sclerosis (AML,Lou Gehrig's disease), multiple sclerosis (MS), schizophrenia,cardiomyocyte hypertrophy, reperfusion/ischemia, stroke, and baldness.

The term “JAK-mediated disease”, as used herein means any disease orother deleterious condition in which a JAK family kinase, in particularJAK-3, is known to play a role. Such conditions include, withoutlimitation, immune responses such as allergic or type I hypersensitivityreactions, asthma, autoimmune diseases such as transplant rejection,graft versus host disease, rheumatoid arthritis, amyotrophic lateralsclerosis, and multiple sclerosis, neurodegenerative disorders such asFamilial amyotrophic lateral sclerosis (FALS), as well as in solid andhematologic malignancies such as leukemias and lymphomas.

The terms “Lck-mediated disease” or “Lck-mediated condition”, as usedherein, mean any disease state or other deleterious condition in whichLck is known to play a role. The terms “Lck-mediated disease” or“Lck-mediated condition” also mean those diseases or conditions that arealleviated by treatment with an Lck inhibitor. Lck-mediated diseases orconditions include, but are not limited to, autoimmune diseases such astransplant rejection, allergies, rheumatoid arthritis, and leukemia. Theassociation of Lck with various diseases has been described [Molina etal., Nature, 1992, 357, 161].

The terms “Src-mediated disease” or “Src-mediated condition”, as usedherein mean any disease or other deleterious condition in which one ormore Src-family kinases is known to play a role. The terms “Src-mediateddisease” or “Src-mediated condition” also mean those diseases orconditions that are alleviated by treatment with an inhibitor of a Srcfamily kinase. Such diseases or conditions include hypercalcemia,restenosis, osteoporosis, osteoarthritis, symptomatic treatment of bonemetastasis, rheumatoid arthritis, inflammatory bowel disease, multiplesclerosis, psoriasis, lupus, graft vs. host disease, T-cell mediatedhypersensitivity disease, Hashimoto's thyroiditis, Guillain-Barresyndrome, chronic obstructive pulmonary disorder, contact dermatitis,cancer, Paget's disease, asthma, ischemic or reperfusion injury,allergic disease, atopic dermatitis, and allergic rhinitis. Src proteinkinase and its implication in various diseases has been described[Soriano, Cell, 1992, 69, 551; Soriano et al., Cell 1991, 64, 693;Takayanagi, J. Clin. Invest. 1999, 104, 137; Boschelli, Drugs of theFuture 2000, 25 (7), 717; Talamonti, J. Clin. Invest. 1993, 91, 53;Lutz, Biochem. Biophys. Res. 1998, 243, 503; Rosen, J. Biol. Chem.,1986, 261, 13754; Bolen, Proc. Natl. Acad. Sci. USA 1987, 84, 2251;Masaki, Hepatology 1998, 27, 1257; Biscardi, Adv. Cancer Res. 1999, 76,61; Lynch, Leukemia 1993, 7, 1416; Wiener, Clin. Cancer Res. 1999, 5,2164; Staley, Cell Growth Diff, 1997, 8, 269]. Diseases that areaffected by Src activity, in particular, include hypercalcemia,osteoporosis, osteoarthritis, cancer, symptomatic treatment of bonemetastasis, and Paget's disease.

The term “SYK-mediated disease” or “SYK-mediated condition”, as usedherein, means any disease or other deleterious condition in which SYKprotein kinase is known to play a role. Such conditions include, withoutlimitation, allergic disorders, especially asthma.

The term “Tec family tyrosine kinases-mediated condition”, as usedherein means any disease or other deleterious condition in which Tecfamily kinases are known to play a role. Such conditions include,without limitation, autoimmune, inflammatory, proliferative, andhyperproliferative diseases and immunologically-mediated diseasesincluding rejection of transplanted organs or tissues and AcquiredImmunodeficiency Syndrome (AIDS).

For example, Tec family tyrosine kinases-mediated conditions includediseases of the respiratory tract including, without limitation,reversible obstructive airways diseases including asthma, such asbronchial, allergic, intrinsic, extrinsic and dust asthma, particularlychronic or inveterate asthma (e.g. late asthma airwayshyper-responsiveness) and bronchitis. Additionally, Tec family tyrosinekinases diseases include, without limitation, those conditionscharacterised by inflammation of the nasal mucus membrane, includingacute rhinitis, allergic, atrophic thinitis and chronic rhinitisincluding rhinitis caseosa, hypertrophic rhinitis, rhinitis purulenta,rhinitis sicca and rhinitis medicamentosa; membranous rhinitis includingcroupous, fibrinous and pseudomembranous rhinitis and scrofoulousrhinitis, seasonal rhinitis including rhinitis nervosa (hay fever) andvasomotor rhinitis, sarcoidosis, farmer's lung and related diseases,fibroid lung and idiopathic interstitial pneumonia.

Tec family tyrosine kinases-mediated conditions also include diseases ofthe bone and joints including, without limitation, (pannus formation in)rheumatoid arthritis, seronegative spondyloarthropathis (includingankylosing spondylitis, psoriatic arthritis and Reiter's disease),Behcet's disease, Sjogren's syndrome, and systemic sclerosis.

Tec family kinases-mediated conditions also include diseases anddisorders of the skin, including, without limitation, psoriasis,systemic sclerosis, atopical dermatitis, contact dermatitis and othereczematous dermatitis, seborrhoetic dermatitis, Lichen planus,Pemphigus, bullous Pemphigus, epidermolysis bullosa, urticaria,angiodermas, vasculitides, erythemas, cutaneous eosinophilias, uveitis,Alopecia, areata and vernal conjunctivitis.

Tec family tyrosine kinases-mediated conditions also include diseasesand disorders of the gastrointestinal tract, including, withoutlimitation, Coeliac disease, proctitis, eosinophilic gastro-enteritis,mastocytosis, pancreatitis, Crohn's disease, ulcerative colitis,food-related allergies which have effects remote from the gut, e.g.migraine, rhinitis and eczema.

Tec family tyrosine kinases-mediated conditions also include thosediseases and disorders of other tissues and systemic disease, including,without limitation, multiple sclerosis, artherosclerosis, acquiredimmunodeficiency syndrome (AIDS), lupus erythematosus, systemic lupus,erythematosus, Hashimoto's thyroiditis, myasthenia gravis, type Idiabetes, nephrotic syndrome, eosinophilia fascitis, hyper IgE syndrome,lepromatous leprosy, sezary syndrome and idiopathic thrombocytopeniapurpura, restenosis following angioplasty, tumours (for exampleleukemia, lymphomas), artherosclerosis, and systemic lupuserythematosus.

Tec family tyrosine kinases-mediated conditions also include allograftrejection including, without limitation, acute and chronic allograftrejection following for example transplantation of kidney, heart, liver,lung, bone marrow, skin and cornea; and chronic graft versus hostdisease.

In other embodiments, the invention relates to a method of enhancingglycogen synthesis and/or lowering blood levels of glucose in a patientin need thereof, comprising administering to said patient atherapeutically effective amount of a composition comprising a compoundof formula I. This method is especially useful for diabetic patients.

In yet another embodiment, the invention relates to a method ofinhibiting the production of hyperphosphorylated Tau protein in apatient in need thereof, comprising administering to said patient atherapeutically effective amount of a composition comprising a compoundof formula I. This method is especially useful in halting or slowing theprogression of Alzheimer's disease.

In still another embodiments, the invention relates to a method ofinhibiting the phosphorylation of β-catenin in a patient in needthereof, comprising administering to said patient a therapeuticallyeffective amount of a composition comprising a compound of formula I.This method is especially useful for treating schizophrenia.

It will also be appreciated that the compounds and pharmaceuticallyacceptable compositions of the present invention can be employed incombination therapies, that is, the compounds and pharmaceuticallyacceptable compositions can be administered concurrently with, prior to,or subsequent to, one or more other desired therapeutics or medicalprocedures. The particular combination of therapies (therapeutics orprocedures) to employ in a combination regimen will take into accountcompatibility of the desired therapeutics and/or procedures and thedesired therapeutic effect to be achieved. It will also be appreciatedthat the therapies employed may achieve a desired effect for the samedisorder (for example, an inventive compound may be administeredconcurrently with another agent used to treat the same disorder), orthey may achieve different effects (e.g., control of any adverseeffects). As used herein, additional therapeutic agents that arenormally administered to treat or prevent a particular disease, orcondition, are known as “appropriate for the disease, or condition,being treated”.

For example, chemotherapeutic agents or other anti-proliferative agentsmay be combined with the compounds of this invention to treatproliferative diseases and cancer. Examples of known chemotherapeuticagents include, but are not limited to, For example, other therapies oranticancer agents that may be used in combination with the inventiveanticancer agents of the present invention include surgery, radiotherapy(in but a few examples, gamma.-radiation, neutron beam radiotherapy,electron beam radiotherapy, proton therapy, brachytherapy, and systemicradioactive isotopes, to name a few), endocrine therapy, biologicresponse modifiers (interferons, interleukins, and tumor necrosis factor(TNF) to name a few), hyperthermia and cryotherapy, agents to attenuateany adverse effects (e.g., antiemetics), and other approvedchemotherapeutic drugs, including, but not limited to, alkylating drugs(mechlorethamine, chlorambucil, Cyclophosphamide, Melphalan,Ifosfamide), antimetabolites (Methotrexate), purine antagonists andpyrimidine antagonists (6-Mercaptopurine, 5-Fluorouracil, Cytarabile,Gemcitabine), spindle poisons (Vinblastine, Vincristine, Vinorelbine,Paclitaxel), podophyllotoxins (Etoposide, Irinotecan, Topotecan),antibiotics (Doxorubicin, Bleomycin, Mitomycin), nitrosoureas(Carmustine, Lomustine), inorganic ions (Cisplatin, Carboplatin),enzymes (Asparaginase), and hormones (Tamoxifen, Leuprolide, Flutamide,and Megestrol), Gleevec™, adriamycin, dexamethasone, andcyclophosphamide. For a more comprehensive discussion of updated cancertherapies see, http://www.nci.nih.gov/, a list of the FDA approvedoncology drugs at http://www.fda.gov/cder/cancer/druglistframe.htm, andThe Merck Manual, Seventeenth Ed. 1999, the entire contents of which arehereby incorporated by reference.

Other examples of agents the inhibitors of this invention may also becombined with include, without limitation: treatments for Alzheimer'sDisease such as Aricept® and Excelon®; treatments for Parkinson'sDisease such as L-DOPA/carbidopa, entacapone, ropinrole, pramipexole,bromocriptine, pergolide, trihexephendyl, and amantadine; agents fortreating Multiple Sclerosis (MS) such as beta interferon (e.g., Avonex®and Rebif®), Copaxone®, and mitoxantrone; treatments for asthma such asalbuterol and Singulair®; agents for treating schizophrenia such aszyprexa, risperdal, seroquel, and haloperidol; anti-inflammatory agentssuch as corticosteroids, TNF blockers, IL-1 RA, azathioprine,cyclophosphamide, and sulfasalazine; immunomodulatory andimmunosuppressive agents such as cyclosporin, tacrolimus, rapamycin,mycophenolate mofetil, interferons, corticosteroids, cyclophosphamide,azathioprine, and sulfasalazine; neurotrophic factors such asacetylcholinesterase inhibitors, MAO inhibitors, interferons,anti-convulsants, ion channel blockers, riluzole, and anti-Parkinsonianagents; agents for treating cardiovascular disease such asbeta-blockers, ACE inhibitors, diuretics, nitrates, calcium channelblockers, and statins; agents for treating liver disease such ascorticosteroids, cholestyramine, interferons, and anti-viral agents;agents for treating blood disorders such as corticosteroids,anti-leukemic agents, and growth factors; and agents for treatingimmunodeficiency disorders such as gamma globulin.

The amount of additional therapeutic agent present in the compositionsof this invention will be no more than the amount that would normally beadministered in a composition comprising that therapeutic agent as theonly active agent. Preferably the amount of additional therapeutic agentin the presently disclosed compositions will range from about 50% to100% of the amount normally present in a composition comprising thatagent as the only therapeutically active agent.

The compounds of this invention or pharmaceutically acceptablecompositions thereof may also be incorporated into compositions forcoating implantable medical devices, such as prostheses, artificialvalves, vascular grafts, stents and catheters. Accordingly, the presentinvention, in another aspect, includes a composition for coating animplantable device comprising a compound of the present invention asdescribed generally above, and in classes and subclasses herein, and acarrier suitable for coating said implantable device. In still anotheraspect, the present invention includes an implantable device coated witha composition comprising a compound of the present invention asdescribed generally above, and in classes and subclasses herein, and acarrier suitable for coating said implantable device.

Vascular stents, for example, have been used to overcome restenosis(re-narrowing of the vessel wall after injury). However, patients usingstents or other implantable devices risk clot formation or plateletactivation. These unwanted effects may be prevented or mitigated bypre-coating the device with a pharmaceutically acceptable compositioncomprising a kinase inhibitor. Suitable coatings and the generalpreparation of coated implantable devices are described in U.S. Pat.Nos. 6,099,562; 5,886,026; and 5,304,121. The coatings are typicallybiocompatible polymeric materials such as a hydrogel polymer,polymethyldisiloxane, polycaprolactone, polyethylene glycol, polylacticacid, ethylene vinyl acetate, and mixtures thereof. The coatings mayoptionally be further covered by a suitable topcoat of fluorosilicone,polysaccarides, polyethylene glycol, phospholipids or combinationsthereof to impart controlled release characteristics in the composition.

Another aspect of the invention relates to inhibiting GSK-3, SYK,Aurora-2, CDK-2, JAK-3, LCK, SRC, and/or Tec family (e.g., Tec, Btk,Itk/Emt/Tsk, Bmx, Txk/Rlk) activity in a biological sample or a patient,which method comprises administering to the patient, or contacting saidbiological sample with a compound of formula I or a compositioncomprising said compound. The term “biological sample”, as used herein,includes, without limitation, cell cultures or extracts thereof;biopsied material obtained from a mammal or extracts thereof; and blood,saliva, urine, feces, semen, tears, or other body fluids or extractsthereof.

Inhibition of GSK-3, SYK, Aurora-2, CDK-2, JAK-3, LCK, SRC, and/or Tecfamily (e.g., Tec, Btk, Itk/Emt/Tsk, Bmx, Txk/Rlk) kinase activity in abiological sample is useful for a variety of purposes that are known toone of skill in the art. Examples of such purposes include, but are notlimited to, blood transfusion, organ-transplantation, biologicalspecimen storage, and biological assays.

SYNTHETIC EXAMPLES

As used herein, The term “Rt(min)” refers to the HPLC retention time, inminutes, associated with the compound. Unless otherwise indicated, theHPLC method utilized to obtain the reported retention time is asfollows:

Column: Ace 5 C8, 15 cm×4.6 mm id

Gradient: 0-100% acetonitrile+methanol (50:50) (20 mM Tris phosphate atpH 7.0)

Flow rate: 1.5 ml/min

Detection: 225 nm

Example 1

2-Bromo-6,7-dihydro-5H-benzo[b]thiophen-4-one

To a solution of 6,7-Dihydro-5H-benzo[b]thiophen-4-one (10.0 g, 65.8mmol) in 50% acetic acid (100 mL) at −5 deg C. was added dropwise asolution of bromine (3.4 mL, 65.8 mmol) in acetic acid (61.5 mL). Themixture was stirred at −5 deg C. for 1 hour.

A solution of 1M NaOAc_((aq)) was added and the resulting precipitateremoved by filtration. This was further washed with water to give thetitled compound as a grey powder (10.8 g). MS (ES⁺) 232. δH (d⁶ DMSO)2.10 (2H, m), 2.47 (2H, t), 2.96 (2H, t) and 7.33 (1H, s).

Example 2

3-(8-Bromo-5,6-dihydro-thieno[2,3-h]quinazolin-2-ylamino)-benzenesulfonamide

To the 2-Bromo-6,7-dihydro-5H-benzo[b]thiophen-4-one (7.8 g, 34.2 mmol)in anhydrous toluene (10 mL) was added DMF/DMA solution (9.2 mL, 68.4mmol) and the mixture was refluxed under nitrogen for 18 hours. Thereaction mixture was concentrated in vacuo and the resulting crude deepred product(2-Bromo-5-dimethylaminomethylene-6,7-dihydro-5H-benzo[b]thiophen-4-one)was taken through to the next step without further purification.

The above was dissolved in anhydrous propan-2-ol (30 mL) and treatedwith 3-Guanidino-benzenesulfonamide hydrogen chloride (10.5 g, 42.0mmol) and powdered sodium hydroxide (1.5 g, 37.6 mmol). The mixture wasrefluxed with vigorous stirring for 5 hours. A yellow precipitate formedduring the reaction and further alcohol was added to maintain theslurry.

The mixture was cooled to room temperature and concentrated in vacuo.The resulting solid was slurried in hot EtOAc (500 mL) and after coolingthe dark green solid product (6.5 g) was removed by filtration. MS (ES⁺)439, (ES⁻) 437. δH (d⁶ DMSO) 2.91 (2H, m), 3.00 (2H, m), 7.30 (2H, brm), 7.37 (1H, m), 7.46 (2H, m), 7.70 (1H, s), 8.39 (1H, s), 8.80 (1H, s)and 9.91 (1H, s).

Example 3

3-(8-Phenyl-5,6-dihydro-thieno[2,3-h]quinazolin-2-ylamino)-benzenesulfonamide

To the3-(8-Bromo-5,6-dihydro-thieno[2,3-h]quinazolin-2-ylamino)-benzenesulfonamide(200 mg, 0.46 mmol) in a 10 mL microwave reaction tube was added Phenylboronic acid (56 mg, 0.46 mmol), 2M Na₂CO_(3(aq)) (920 μL, 1.84 mmol),Pd(PPh₃)₄ (5.3 mg, 0.0046 mmol), 3:4 EtOH/H₂O (1.5 mL) and DME (2.5 mL).

The tube was capped and subjected to microwave irradiation at 120 deg C.for 2 hours. On cooling the organic layer was separated from thereaction mixture and purified by preparative liquid chromatography. Thedesired compound (67 mg) was obtained as a yellow powder following thefreeze-drying of product fractions. MS (ES⁺) 435, (ES⁻) 433. δH (d⁶DMSO) 2.94 (2H, m), 3.05 (2H, m), 7.30 (3H, m), 7.40 (4H, m), 7.60 (1H,d), 7.80 (2H, d), 8.01 (1H, s), 8.40 (1H, s), 9.10 (1H, s) and 9.90 (1H,s).

A variety of other compounds of formula I have been prepared by methodssubstantially similar to those described in Example 3. Thecharacterization data for these compounds is summarized in Table 9 belowand includes HPLC, LC/MS (observed) and ¹H NMR data.

¹H NMR data is summarized in Table 9 below wherein ¹H NMR data wasobtained at 400 MHz in deuterated DMSO, unless otherwise indicated, andwas found to be consistent with structure. Compound numbers correspondto the compound numbers listed in Table 1.

TABLE 9 Characterization Data for Selected Compounds of Formula ICompound M + 1 Rt No. (obs) (min) ¹H-NMR I-2 465 9.92 2.95 (2H, m), 3.07(2H, m), 3.90 (3H, s), 6.90 (1H, d), 7.31 (5H, m), 7.40 (1H, m), 7.46(1H, t), 7.69 (1H, d), 8.06 (1H, s), 8.40 (1H, s), 9.20 (1H, s), 9.95 ((1H, s). I-3 436 8.84 2.98 (2H, m), 3.12 (2H, m), 6.90 (1H, d), 7.35(2H, s), 7.40 (1H, d), 7.48 (1H, t), 7.60 (2H, m), 8.20 (1H, s), 8.29(1H, d), 8.41 (1H, s), 8.59 (1H, m), 8.85 (1H, d), 9.90 (1H, s), 9.98 ((1H, s). I-4 479 7.46 2.98 (2H, m), 3.09 (2H, m), 7.30 (1H, t), 7.39(1H, s), 7.48 (3H, m), 7.77 (2H, d), 7.80 (1H, d), 8.00 (1H, s), 8.19(1H, s), 8.40 (1H, s), 8.90 (1H, br s), 9.95 ( (1H, s). I-5 441 9.722.97 (2H, m), 3.07 (2H, m), 7.35 (2H, s), 7.40 (1H, d), 7.48 (1H, t),7.61 (2H, m), 7.70 (1H, m), 7.81 (1H, s), 8.93 (1H, s), 8.39 (1H, s),9.18 (1H, s), 9.91 (1H, s). I-6 492 8.93 2.07 (3H, s), 2.97 (2H, m),3.11 (2H, m), 7.32 (2H, s), 7.39 (2H, m), 7.49 (2H, d), 7.51 (1H, m),7.70 (1H, m), 7.98 (1H, s), 8.03 (1H, s), 8.39 (1H, s), 8.89 (1H, s),9.93 (1H, s), 10.03 (1H, s). I-7 469 10.12 2.98 (2H, m), 3.09 (2H, m),7.29 (2H, s), 7.50 (4H, br m), 7.60 (1H, d), 7.65 (2H, m), 7.96 (1H, s),8.40 (1H, s), 8.79 (1H, s), 9.90 (1H, s). I-8 441 9.67 2.95 (2H, m),3.17 (2H, m), 7.10 (1H, m), 7.22 (2H, s), 7.28 (1H, d), 7.34 (2H, m),7.55 (2H, d), 7.68 (1H, d), 7.85 (1H, s), 8.40 (1H, s), 8.95 (1H, s),9.91 (1H, s). I-9 465 9.72 2.95 (2H, m), 3.07 (2H, m), 3.95 (3H, s),7.11 (1H, t), 7.15 (1H, d), 7.29 (2H, s), 7.31 (1H, m), 7.40 (1H, d),7.48 (1H, t), 7.70 (1H, d), 7.91 (1H, d), 8.09 (1H, s), 8.38 (1H, s),8.91 (1H, s), 9.86 (1H, s). I-10 450 8.93 2.95 (2H, m), 3.09 (2H, m),6.81 (1H, m), 7.20 (1H, s), 7.25 (1H, m), 7.34 (3H, s), 7.35 (1H, d),7.44 (1H, m), 7.68 (1H, m), 7.95 (1H, s), 8.40 (1H, s), 8.98 (1H, s),8.91 (1H, s). I-11 465 9.78 2.92 (2H, m), 3.05 (2H, m), 3.80 (3H, s),6.95 (2H, d), 7.31 (1H, m), 7.39 (1H, d), 7.46 (1H, t), 7.48 (1H, t),7.61 (1H, m), 7.70 (2H, d), 7.99 (1H, s), 8.39 (1H, s), 9.06 (1H, br s),9.89 (1H, s). I-12 460 9.38 2.98 (2H, m), 3.10 (2H, m), 7.20 (2H, br s),7.40 (1H, m), 7.43 (1H, m), 7.60 (2H, br m), 7.80 (1H, d), 8.09 (1H, d),8.20 (1H, s), 8.26 (1H, s), 8.41 (1H, s), 9.15 (1H, s), 9.93 (1H, s).I-13 480 9.67 2.98 (2H, m), 3.11 (2H, m), 7.31 (2H, s), 7.38 (1H, d),7.47 (1H, t), 7.62 (1H, d), 7.83 (1H, t), 8.20 (2H, br m), 8.30 (1H, brm), 8.41 (2H, m), 9.09 (1H, s), 9.95 (1H, s). I-14 495 9.52 2.95 (2H,m), 3.05 (2H, m), 6.80 (2H, d), 7.28 (2H, s), 7.31 (1H, t), 7.39 (1H,m), 7.45 (1H, t), 7.81 (1H, s), 7.99 (1H, s), 8.39 (1H, s), 8.46 (1H,s), 9.80 (1H, s). I-15 469 10.12 2.96 (2H, m), 3.09 (2H, m), 7.27 (2H,s), 7.39 (2H, m), 7.48 (2H, t), 7.61 (1H, d), 7.75 (1H, d), 7.90 (1H,s), 8.11 (1H, s), 8.40 (1H, s), 9.15 (1H, s), 9.90 (1H, s). I-16 4699.99 2.95 (2H, m), 3.15 (2H, m), 7.31 (2H, s), 7.40 (1H, d), 7.45 (3H,m), 7.63 (1H, d), 7.79 (1H, d), 8.08 (1H, s), 8.40 (1H, s), 9.06 (1H,s), 9.89 (1H, s). I-17 495 9.20 2.98 (2H, m), 3.09 (2H, m), 3.81 (3H,s), 3.87 (3H, s), 7.01 (1H, d), 7.15 (1H, t), 7.31 (1H, d), 7.481H, t),7.60 (1H, d), 7.70 (1H, d), 8.09 (1H, s), 8.39 (1H, s), 8.91 (1H, s) and9.83 (1H, s). I-20 495 9.20 2.92 (2H, m), 3.05 (2H, m), 3.85 (3H, s),3.87 (3H, s), 6.88 (1H, m), 7.09 (1H, d), 7.27 (1H, br s), 7.37 (1H, m),7.44 (1H, t), 7.70 (1H, m), 8.07 (1H, s), 8.39 (1H, s), 8.98 (1H, br s)and 9.82 (1H, s). I-26 528 8.97 2.99 (2H, m), 3.10 (2H, m), 6.88 (1H,m), 7.17 (1H, d), 7.30 (2H, br s), 7.40 (1H, d), 7.46 (1H, m), 7.50 (1H,m), 7.59 (1H, d), 7.71 (1H, d), 7.99 (1H, s), 8.40 (1H, s), 8.90 (1H,s), 9.81 (1H, s) and 9.89 (1H, s). I-27 451 8.97 2.95 (2H, m), 3.05 (2H,m), 6.85 (1H, t), 6.95 (1H, m), 7.13 (1H, t), 7.29 (2H, br s), 7.40 (1H,d), 7.46 (1H, t), 7.78 (1H, d), 7.81 (1H, d), 8.09 (1H, s), 8.36 (1H,s), 8.83 (1H, s), 9.83 (1H, s) and 10.29 (1H, br s). I-28 519 10.28 2.99(2H, m), 3.13 (2H, m), 7.29 (2H, br s), 7.36 (1H, d), 7.40-7.51 (4H, brm), 7.75 (1H, d), 8.06 (2H, s), 8.42 (1H, s), 8.86 (1H, s) and 10.29(1H, br s). I-29 465 9.12 2.97 (2H, m), 3.09 (2H, m), 4.63 (2H, d), 5.26(1H, m), 7.23 (2H, br s), 7.37 (2H, m), 7.43 (3H, br m), 7.63 (1H, d),7.75 (1H, s), 7.91 (1H, d), 8.39 (1H, s), 8.60 (1H, s) and 9.82 (1H, s).

Example 4

2-(3-Methoxyphenyl)-6,7-dihydro-5H-benzo[b]thiophen-4-one:

To 2-bromo-6,7-dihydro-5H-benzo[b]thiophen-4-one (5 g, 21.74 mmol) wassuccessively added 3-methoxyphenol boronic acid (3.63 g, 23.92 mmol), 2MNa₂CO_(3(aq)) (43.5 mL, 86.98 mmol), Pd(PPh₃)₄ (251 mg, 0.22 mmol), 3:4EtOH/H₂O (37.5 mL) and DME (62.5 mL). The reaction mixture was degassedand heated to reflux for 12 hours. EtOAc (250 mL) was added and thecrude reaction mixture was washed with water (2×100 mL). The organiclayer was dried over magnesium sulphate, filtered and concentrated invacuo. The residue was purified by silica gel chromatography elutingwith EtOAc:hexanes (20:80) to give the title compound as an off-whitesolid in 95% yield. MS (ES⁺) 259. δH (d⁶ DMSO) 2.13 (2H, quint.), 2.50(2H, m), 3.05 (2H, t), 3.81 (3H, s), 6.91 (1H, m), 7.20-7.22 (2H, m),7.34 (1H, m), 7.67 (1H, s).

Example 5

4-[8-(3-Methoxyphenyl)-5,6-dihydro-thieno[2,3-h]quinazolin-2-ylamino)-benzenesulfonamide

To 2-(3-methoxyphenyl)-6,7-dihydro-5H-benzo[b]thiophen-4-one (0.3 g,1.16 mmol) was added DMF/DMA solution (2 mL) and the mixture wasrefluxed under nitrogen for 12 hours. The reaction mixture wasconcentrated in vacuo and the resulting crude product(5-dimethylaminomethylene-2-(3-methoxyphenyl)-6,7-dihydro-5H-benzo[b]thiophen-4-one)was taken through to the next step without further purification.

The above was dissolved in anhydrous DMA (3 mL) and treated with4-Guanidino-benzenesulfonamide hydrogen chloride (297 mg, 1.18 mmol) andpowdered potassium carbonate (82 mg, 0.59 mmol). The mixture was heatedto 120° C. with vigorous stirring for 12 hours. The mixture was cooleddown to room temperature and purified by reverse phase preparative HPLC[Waters Delta-Pak C18, 15 uM, 100 A column, gradient 10%-100% B (solventA: 0.05% TFA in water; solvent B: CH₃CN) over 10 minutes at 25 mL/min]to afford the title compound (17 mg) as a yellow powder. MS (ES⁺) 465,(ES⁻) 463. δH (d⁶ DMSO) 2.99 (2H, t), 3.10 (2H, t), 3.85 (3H, s), 6.95(1H, dd), 7.13 (2H, br s), 7.21 (1H, m), 7.27 (1H, d), 7.40 (1H, t),7.76 (2H, d), 7.87 (1H, s), 8.01 (2H, d), 8.42 (1H, s), 9.90 (1H, s).

Other compounds of formula I have been prepared by methods substantiallysimilar to those described in Example 5. The characterization data forthese compounds is summarized in Table 10 below and includes HPLC, LC/MS(observed) and ¹H NMR data.

¹H NMR data is summarized in Table 10 below wherein ¹H NMR data wasobtained at 400 MHz in deuterated DMSO, unless otherwise indicated, andwas found to be consistent with structure. Compound numbers correspondto the compound numbers listed in Table 1.

TABLE 10 Characterization Data for Selected Compounds of Formula ICompound M + 1 Rt No. (obs) (min) ¹H-NMR I-19 402 10.23 2.96 (2H, t),3.08 (2H, t), 3.84 (3H, s), 6.36 (1H, dd), 6.94 (1H, dd), 7.06 (1H, t),7.14 (1H, d), 7.21 (1H, t), 7.24 (1H, d), 7.38 (1H, t), 7.50 (1H, t),7.87 (1H, s), 8.33 (1H, s), 9.37 (1H, s) I-22 386 10.94 2.96 (2H, t),3.08 (2H, t), 3.84 (3H, s), 6.90-6.97 (2H, m), 7.19 (1H, t), 7.24 (1H,d), 7.30 (2H, t), 7.39 (1H, t), 7.80-7.86 (3H, m), 8.35 (1H, s), 9.44(1H, s) I-23 402 10.14 2.93 (2H, t), 3.07 (2H, t), 3.84 (3H, s), 6.74(2H, d), 6.94 (1H, dd), 7.18 (1H, t), 7.23 (1H, d), 7.38 (1H, t), 7.54(2H, d), 7.78 (1H, s), 8.25 (1H, s), 9.27 (1H, br s) I-24 464 10.302.97-3.02 (2H, m), 3.05-3.10 (2H, m), 3.22 (3H, s), 3.88 (3H, s), 6.91(1H, d), 7.27-7.37 (3H, m), 7.45 (1H, d), 7.55 (1H, t), 7.70 (1H, d),8.02 (1H, s), 8.41 (1H, s), 9.21 (1H, s), 10.00 (1H, s) I-25 464 10.302.96-3.03 (2H, m), 3.05-3.15 (2H, m), 3.15 (3H, s), 3.85 (3H, s), 6.96(1H, d), 7.20, (1H, s), 7.26 (1H, d), 7.40 (1H, t), 7.83-7.90 (3H, m),8.10 (2H, d), 8.41 (1H, s), 10.05 (1H, s) I-35 429 10.04 2.97 (2H, t),3.09 (2H, t), 3.88 (3H, s), 6.93 (1H, dd), 7.21 (1H, br s), 7.26 (1H,t), 7.30-7.40 (3H, m), 7.44 (1H, d), 7.72 (1H, dd), 7.89 (1H, br s),8.02 (1H, s), 8.37 (1H, s), 8.82 (1H, s), 9.62 (1H, s) I-36 443 10.042.13 (3H, s), 2.96 (2H, t), 3.07 (2H, t), 3.83 (3H, s), 6.92-6.96 (2H,m), 7.14-7.17 (2H, m), 7.34-7.43 (3H, m), 8.27 (1H, s), 8.33 (1H, s),8.89 (1H, s), 9.44 (1H, s), 9.90 (1H, s) I-37 493 10.62 2.63 (6H, s),2.99 (2H, t), 3.10 (2H, t), 3.87 (3H, s), 6.93 (1H, d), 7.26- 7.39 (4H,m), 7.55 (1H, t), 7.74 (1H, dd), 8.01 (1H, s), 8.42 (1H, s), 9.14 (1H,s), 9.99 (1H, s) I-38 479 10.42 2.46 (3H, d), 2.99 (2H, t), 3.10 (2H,t), 3.88 (3H, s), 6.92 (1H, dd), 7.28- 7.38 (5H, m), 7.50 (1H, t), 7.66(1H, dd), 8.04 (1H, s), 8.41 (1H, s), 9.16 (1H, s), 9.92 (1H, s) I-39479 15.58 2.93-3.10 (7H, m), 3.85 (3H, s), 6.76 (1H, d), 6.93 (1H, dd),7.19-7.45 (5H, m), 8.01 (1H, s), 8.14 (1H, s), 8.35 (1H, s), 9.52 (1H,s), 9.67 (1H, br s) I-40 401 10.29 2.94 (2H, t), 3.07 (2H, t), 3.84 (3H,s), 4.96 (2H, s), 6.17 (1H, m), 6.88- 6.95 (3H, m), 7.22-7.26 (3H, m),7.37 (1H, t), 7.87 (1H, s), 8.30 (1H, s), 9.11 (1H, s) I-46 402 10.652.97 (2H, t), 3.09 (2H, t), 3.85 (3H, s), 6.80-6.96 (4H, m), 7.20-7.28(3H, m), 7.37 (2H, t), 7.88 (1H, s), 8.15- 8.20 (2H, m), 8.34 (1H, s)

Example 6

8-(3-Methoxyphenyl)-5,6-dihydro-thieno[2,3-h]quinazolin-2-ylamine

To 2-(3-methoxyphenyl)-6,7-dihydro-5H-benzo[b]thiophen-4-one (1.78 g,6.89 mmol) was added DMF/DMA solution (12 mL) and the mixture wasrefluxed under nitrogen for 12 hours. The reaction mixture wasconcentrated in vacuo and the resulting crude product(5-dimethylaminomethylene-2-(3-methoxyphenyl)-6,7-dihydro-5H-benzo[b]thiophen-4-one)was taken through to the next step without further purification.

The above was dissolved in anhydrous DMA (18 mL) and treated withguanidine hydrogen chloride (724 mg, 7.58 mmol) and powdered potassiumcarbonate (524 mg, 3.79 mmol). The mixture was heated to 120° C. withvigorous stirring for 12 hours. The mixture was cooled down to roomtemperature. Water was added and the solid was filtered and rinsed withmore to afford the title compound. MS (ES⁺) 310.

Example 7

4-tert-Butyl-N-[8-(3-methoxyphenyl)-(5,6-dihydrothieno[2,3-h]quinazolin-2-yl)-benzamide

4-tert-Butylbenzoyl chloride (0.171 mL, 0.88 mmol) was added to8-(3-methoxyphenyl)-(5,6-dihydrothieno[2,3-h]quinazolin-2-ylamine (0.129g, 0.42 mmol) in pyridine (1 mL). The reaction mixture was stirred atroom temperature overnight. The reaction mixture was concentrated invacuo. The residue was taken up in dichloromethane (20 mL), washed withwater (20 mL×2) and saturated sodium bicarbonate solution (20 mL×2),dried (MgSO₄) and concentrated in vacuo. The resulting crude mixture waspurified by silica gel chromatography to afford the title compound as abeige solid (75 mg, 30% yield). MS (ES⁺) 470, (ES⁻) 468. δH (d⁶ DMSO)1.32 (9H, s), 3.02-3.15 (4H, m), 3.83 (3H, s), 6.93 (1H, dd), 7.17 (1H,t), 7.22 (1H, d), 7.36 (1H, t), 7.54 (2H, d), 7.78 (1H, s), 7.94 (2H,d), 8.56 (1H, s), 10.74 (1H, s).

A variety of other compounds of formula I have been prepared by methodssubstantially similar to those described in Example 7. Thecharacterization data for these compounds is summarized in Table 11below and includes HPLC, LC/MS (observed) and ¹H NMR data.

¹H NMR data is summarized in Table 11 below wherein ¹H NMR data wasobtained at 400 MHz in deuterated DMSO, unless otherwise indicated, andwas found to be consistent with structure. Compound numbers correspondto the compound numbers listed in Table 1.

TABLE 11 Characterization Data for Selected Compounds of Formula ICompound M + 1 Rt No. (obs) (min) ¹H-NMR I-30 378 9.71 0.78-0.87 (4H,m), 2.14-2.21 (1H, m), 2.98-3.12 (4H, m), 3.84 (3H, s), 6.94 (1H, dd),7.17 (1H, t), 7.22 (1H, d), 7.37 (1H, t), 7.78 (1H, s), 8.48 (1H, s),10.65 (1H, s) I-32 414 9.96 3.03-3.16 (4H, m), 3.83 (3H, s), 6.93 (1H,dd), 7.17 (1H, t), 7.23 (1H, d), 7.37 (1H, t), 7.52 (2H, t), 7.61 (1H,t), 7.76 (1H, s), 7.98 (2H, d), 8.57 (1H, s), 10.84 (1H, s) I-33 45710.18 3.02 (6H, s), 3.02-3.15 (4H, m), 3.83 (3H, s), 6.75 (2H, d), 6.93(1H, dd), 7.18 (1H, t), 7.23 (1H, d), 7.36 (1H, t), 7.81 (1H, s), 7.90(2H, d), 8.54 (1H, s), 10.40 (1H, s) I-34 403 9.72 3.02-3.15 (4H, m),3.84 (3H, s), 6.17 (1H, m), 6.93 (1H, dd), 7.00 (1H, m), 7.18-7.20 (2H,m), 7.23 (1H, d), 7.37 (1H, t), 7.85 (1H, s), 8.54 (1H, s), 10.35 (1H,s), 11.68 (1H, br s)

Example 8

3-(8-Phenyl-thieno[2,3-h]quinazolin-2-ylamino)-benzenesulfonamide

To a dark solution of3-(8-Phenyl-5,6-dihydro-thieno[2,3-h]quinazolin-2-ylamino)-benzenesulfonamide(130 mg, 0.30 mmol) in anhydrous 1,4-dioxane (7 mL) was added DDQ (75mg, 0.33 mmol) and the mixture was refluxed under nitrogen for 2-3hours. The reaction mixture was concentrated in vacuo and the resultingcrude residue was triturated in a mixture of 2M NaOH (25 mL) and DCM (25mL). The resulting solid was filtered and washed with DCM and water toafford 50 mg (39% yield) of the title compound as a pale yellow/greenpowder. MS (ES⁺) 433, (ES⁻) 431. δH (d⁶ DMSO) 7.39 (2H, br s), 7.41 (1H,m), 7.51 (4H, br m), 7.72 (1H, d), 7.83 (1H, d), 7.96 (3H, m), 8.68 (1H,s), 9.40 (1H, s), 9.61 (1H, br s) and 10.41 (1H, s).

A variety of other compounds of formula II have been prepared by methodssubstantially similar to those described in Example 8. Thecharacterization data for these compounds is summarized in Table 12below and includes HPLC, LC/MS (observed) and ¹H NMR data.

¹H NMR data is summarized in Table 12 below wherein ¹H NMR data wasobtained at 400 MHz in deuterated DMSO, unless otherwise indicated, andwas found to be consistent with structure. Compound numbers correspondto the compound numbers listed in Table 2.

TABLE 12 Characterization Data for Selected Compounds of Formula IICompound M + 1 Rt No. (obs) (min) ¹H-NMR II-31 357 8.55 7.37 (2H, s),7.48 (1H, d), 7.55 (1H, t), 7.84 (1H, d), 7.90 (1H, dd), 7.98- 8.03 (2H,m), 8.29 (1H, d), 9.24 (1H, s), 9.40 (1H, s), 10.39 (1H, s) II-32 30610.52 2.32 (6H, s), 6.67 (1H, s), 7.72 (2H, s), 7.79 (1H, d), 7.95 (1H,d), 7.98 (1H, d), 8.06 (1H, d), 9.33 (1H, s), 9.87 (1H, s) II-33 3358.77 2.11 (3H, s), 7.11 (1H, d), 7.25 (1H, t), 7.53 (1H, d), 7.80 (1H,d), 7.92- 7.97 (2H, m), 8.44 (1H, d), 8.72 (1H, s), 9.34 (1H, s), 9.96(1H, s), 9.99 (1H, s) II-34 293 8.75 6.91 (1H, d), 7.43 (1H, t), 7.73(1H, d), 7.84 (1H, d), 7.98-8.03 (2H, m), 8.32 (1H, d), 8.38 (1H, s),9.40 (1H, s), 10.30 (1H, s) II-35 372 7.60 2.92 (3H, s), 7.37 (2H, s),7.49 (1H, d), 7.56 (1H, t), 7.89 (1H, d), 7.97 (1H, d), 8.09 (1H, d),8.68 (1H, s), 9.44 (1H, s), 10.45 (1H, s). II-36 472 9.0 1.28 (9H, s),5.62 (2H, s), 7.47-7.55 (2H, m), 7.98 (2H, d), 8.04 (1H, d), 8.79 (1H,s), 9.50 (1H, s), 10.50 (1H, s) II-37 443 7.6 3.67 (4H, t), 3.79 (4H,t), 7.35 (2H, s), 7.46 (1H, m), 7.51-7.56 (2H, m), 7.84 (1H, d), 8.12(1H, d), 8.62 (1H, t), 9.27 (1H, s), 10.30 (1H, s) II-38 429 8.5 1.26(6H, t), 3.63 (4H, m), 7.35 (2H, s), 7.40-7.60 (3H, m), 7.78 (1H, d),8.11 (1H, d), 8.63 (1H, s), 9.22 (1H, s), 10.24 (1H, s) II-39 441 8.81.68 (6H, s), 3.69 (4H, s), 7.35 (2H, s), 7.45 (1H, d), 7.50 (1H, d),7.54 (1H, t), 7.80 (1H, d), 8.11 (1H, dd), 8.64 (1H, t), 9.24 (1H, s),10.26 (1H, s)

Example 9

3-(4H-3,5-Dithia-7,9-diazacyclopenta[α]-naphthalen-8-ylamino)-benzenesulfonamide

To 7H-thieno[2,3-c]thiopyran-4-one (495 mg, 2.91 mmol) dissolved inanhydrous DME (5 mL), was added Bredereck's reagent (0.9 mL, 4.36 mmol)and the reaction mixture was heated to 60° C. for 2 hours. The reactionmixture was cooled down and concentrated in vacuo. The resulting residuewas purified by silica gel chromatography eluting with a gradient ofEtOAc:hexanes to give the enaminone as a yellow solid in 58% yield.

The above (100 mg, 0.44 mmol) was dissolved in anhydrous propan-2-ol (5mL) and treated with 3-Guanidino-benzenesulfonamide hydrogen chloride(172 mg, 0.66 mmol) and powdered sodium hydroxide (20 mg, 0.49 mmol).The mixture was refluxed with vigorous stirring for 12 hours. Themixture was cooled down to room temperature and concentrated in vacuo.The resulting residue was purified by silica gel chromatography elutingwith EtOAc:hexanes (50:50) to afford the title compound (25 mg) as ayellow solid. MS (ES⁺) 377. δH (CDCl₃) 4.20 (2H, d), 4.80 (2H, br s),7.20-7.35 (2H, m), 7.45-7.80 (4H, m), 8.40 (1H, s), 8.80 (1H, s).

Example 10

3-(5,6-Dihydrothieno[2,3-h]quinazolin-2-ylamino)-benzenesulfonamide

To 6,7-dihydro-5H-benzo[b]thiophen-4-one (0.5 g, 3.29 mmol) was addedDMF/DMA solution (2 mL) and the mixture was refluxed under nitrogen for12 hours. The reaction mixture was concentrated in vacuo and theresulting crude product(5-dimethylaminomethylene-6,7-dihydro-5H-benzo[b]thiophen-4-one) wastaken through to the next step without further purification.

The above was dissolved in anhydrous DMA (3 mL) and treated with4-guanidino-benzenesulfonamide hydrogen chloride (1.23 g, 4.93 mmol) andpowdered potassium carbonate (341 mg, 2.47 mmol). The mixture was heatedto 120° C. with vigorous stirring for 12 hours. The mixture was cooleddown to room temperature and concentrated in vacuo. The residue wastriturated with hot ethyl acetate (30 ml) and 1M Hcl (30 ml), filteredand washed with water and ethyl acetate to afford the title compound(546 mg) as a pale green powder in 46% yield. MS (ES⁺) 359, (ES⁻) 357.δH (d⁶ DMSO) 2.93 (2H, m), 3.04 (2H, m), 6.88 (1H, m), 7.29 (1H, br s),7.40 (1H, m), 7.45 (1H, t), 7.65 (1H, d), 7.77 (1H, d), 8.40 (1H, s),8.70 (1H, br s) and 10.05 (1H, br s).

A variety of other compounds of formula I have been prepared by methodssubstantially similar to those described in Example 10. Thecharacterization data for these compounds is summarized in Table 13below and includes HPLC, LC/MS (observed) and ¹H NMR data.

¹H NMR data is summarized in Table 13 below wherein ¹H NMR data wasobtained at 400 MHz in deuterated DMSO, unless otherwise indicated, andwas found to be consistent with structure. Compound numbers correspondto the compound numbers listed in Table 1.

TABLE 13 Characterization Data for Selected Compounds of Formula I Com-pound M + 1 Rt No. (obs) (min) ¹H-NMR I-42 325 10.32 2.98 (2H, t), 3.10(2H, t), 7.5-7.6 (2H, m), 7.65 (1H, d), 7.76 (1H, d), 8.04 (1H, d), 8.43(1H, s), 9.20 (1H, s), 10.09 (1H, s) I-43 308 10.59 2.25 (6H, s), 2.91(2H, t), 3.04 (2H, t), 6.57 (1H, s), 7.45-7.54 (4H, m), 8.32 (1H, s),9.32 (1H, s) I-44 337 8.69 2.06 (3H, s), 2.93 (2H, t), 3.05 (2H, t),7.11-7.21 (2H, m), 7.39 (1H, d), 7.49 (1H, d), 7.71 (1H, d), 8.18 (1H,s), 8.31 (1H, s), 9.59 (1H, s), 9.90 (1H, s) I-45 434 8.85 1.40-1.55(6H, m), 2.45-2.67 (8H, m), 2.92 (2H, t), 3.05 (2H, t), 7.14-7.20 (2H,m), 7.35-7.41 (1H, m), 7.47 (1H, d), 7.71 (1H, d), 8.20 (1H, s), 8.31(1H, s), 9.48 (1H, s), 10.23 (1H, s) I-47 366 7.80 2.72 (2H, t), 2.93(2H, t), 3.02-3.14 (4H, m), 7.20 (2H, d), 7.41 (1H, m), 7.48 (1H, d),7.71 (1H, d), 7.76 (3H, br s), 8.21 (1H, s), 8.32 (1H, s), 9.54 (1H, s),10.11 (1H, s) I-48 394 8.33 2.21 (6H, s), 2.46 (2H, t), 2.60 (2H, t),2.92 (2H, t), 3.05 (2H, t), 7.11-7.19 (2H, m), 7.37 (1H, d), 7.47 (1H,d), 7.72 (1H, d), 8.24 (1H, s), 8.32 (1H, s), 9.46 (1H, s), 9.99 (1H, s)I-49 449 8.88 2.14 (3H, s), 2.20-2.52 (10H, t), 2.63 (2H, t), 2.92 (2H,t), 3.05 (2H, t), 7.12-7.20 (2H, m), 7.38 (1H, d), 7.47 (1H, d), 7.72(1H, d), 8.22 (1H, s), 8.32 (1H, s), 9.47 (1H, s), 10.07 (1H, s) I-50436 8.81 2.43 (4H, m), 2.64 (2H, t), 2.92 (2H, t), 3.05 (2H, t), 3.60(4H, t), 7.15-7.20 (2H, m), 7.39 (1H, m), 7.47 (1H, d), 7.71 (1H, d),8.20 (1H, s), 8.31 (1H, s), 9.47 (1H, s), 10.06 (1H, s)

Example 11

2-Methylsulfanyl-5,6-dihydro-thieno[2,3-h]quinazoline-8-carboxylic acidtert-butyl ester

5-[1-Dimethylamino-methylidene]-4-oxo-4,5,6,7-tetrahydro-benzo[b]thiophene-2-carboxylicacid tert-butyl ester (14.36 g, 46.72 mmol), Na₂CO₃ (8.17 g, 77.09 mmol)and S-methylisothiouronium sulphate (19.51 g, 70.08 mmol) weresuspended/dissolved in dry DMA (140 mL) and stirred at 120° C. for 1.5hours. The solvent was removed under reduced pressure and the resultantbrown solid mass was partitioned between hot EtOAc and brine usingsonication. The aqueous layer was further extracted with EtOAc (3×200mL) and the combined organics were washed sequentially with diluteNaHSO₄ (1×100 mL) [It took 15 minutes for the emulsified mixture toseparate fully.], saturated Na₂CO₃ (1×100 mL) and brine (1×100 mL),dried over Na₂SO₄, filtered and concentrated under reduced pressure togive an ochre-brown solid. This solid was redissolved in DCM, silica(˜100 mL) was added and the resulting suspension was concentrated underreduced pressure. The solid obtained was subjected to columnchromatography (gradient elution, 20-40% EtOAc in hexanes, ˜1 L silica)giving a light yellow solid (8.78 g, 56% yield). MS (ES⁺) 335, (ES⁻)332. δH (CDCl₃) 1.6 (9H, s), 2.6 (3H, s), 3.0-3.2 (4H, m), 8.2 (1H, s),8.3 (1H, s)

Example 12

2-Methylsulfanyl-thieno[2,3-h]quinazoline-8-carboxylic acid tert-butylester

2-Methylsulfanyl-5,6-dihydro-thieno[2,3-h]quinazoline-8-carboxylic acidtert-butyl ester (9.0 g, 26.91 mmol) and DDQ (12.22 g, 53.82 mmol) weresuspended/dissolved in dry dioxane (100 mL) and heated at reflux for 1hour. Further DDQ (12.22 g, 53.82 mmol) was added and the resultingsuspension was heated at reflux for 1.5 hours. After cooling to roomtemperature the mixture was concentrated under reduced pressure and thenpartitioned between EtOAc and saturated Na₂CO₃. The aqueous layer wasfurther extracted with EtOAc (3×200 mL) and the combined organics werewashed with saturated Na₂CO₃ (2×250 mL) and brine (1×200 mL), dried overNa₂SO₄, filtered and concentrated under reduced pressure. The deep browngum obtained was redissolved in hot acetone, silica (˜60 mL) was addedand the resultant suspension was concentrated under reduced pressure.The solid obtained was subjected to column chromatography (gradientelution, 20-30% acetone in hexanes, ˜600 mL silica) giving a light brownsolid (6.50 g, 73% yield)). MS (ES⁺) 333. δH (CDCl₃) 1.7 (9H, s), 2.8(3H, s), 7.8 (1H, d), 7.9 (1H, d), 8.8 (1H, s), 9.2 (1H, s)

Example 13

2-Methanesulfonyl-thieno[2,3-h]quinazoline-8-carboxylic acid tert-butylester

2-Methylsulfanyl-thieno[2,3-h]quinazoline-8-carboxylic acid tert-butylester (6.50 g, 19.55 mmol) was suspended/dissolved in DCM (150 mL) andcooled in an ice-bath. 3-chloroperbenzoic acid (22.49 g, 97.76 mmol,˜75% pure) was added in one portion and the resultant suspension wasstirred at 0° C. for 5 minutes and at room temperature for a further 55minutes. This mixture was then carefully added to a 1:1:1 mixture ofsaturated Na₂CO₃, saturated Na₂S₂O₃ and brine (˜500 mL). The organiclayer was separated and the aqueous layer was further extracted with DCM(3×200 mL). The combined organics were washed with brine (1×100 mL),dried over Na₂SO₄, filtered and concentrated under reduced pressure. Theyellow solid obtained was redissolved in acetone, silica (˜80 mL) wasadded and the suspension was concentrated under reduced pressure. Thesolid obtained was subjected to column chromatography (gradient elution,5-10% EtOAc in DCM, ˜600 mL silica) to give a yellow solid (6.25 g, 88%yield). MS (ES⁺) 365. δH (CDCl₃) 1.7 (9H, s), 3.5-3.6 (3H, s), 8.0 (1H,d), 8.2 (1H, d), 8.9 (1H, s), 9.6 (1H, s)

Example 14

2-Methanesulfonyl-thieno[2,3-h]quinazoline-8-carboxylic acid

2-Methanesulfonyl-thieno[2,3-h]quinazoline-8-carboxylic acid tert-butylester (4.20 g, 11.52 mmol) was placed in a 500 mL Florentine and cooledin an ice-bath. Pre-mixed TFA:DCM:water (1:1:0.025, 4.5 mL) was added inone portion and the resultant solution was stirred at 0° C. for 0.75hours and a further 0.75 hours at room temperature. The reaction mixturewas concentrated under reduced pressure and azeotroped with portions ofDCM (5×50 mL) and Et₂O (5×50 mL). The solid obtained was triturated withEt₂O, filtered and washed with Et₂O (3×5 mL) to give a light yellowpowder (3.51 g, 99% yield). δH (d⁶ DMSO) 3.6-3.7 (3H, s), 8.3 (1H, d),8.6 (1H, d), 8.8 (1H, s), 9.9-10.0 (1H, s), 13.8-14.0 (1H, br s)

Example 15

4-(2-Methanesulfonyl-thieno[2,3-h]quinazoline-8-carbonyl)-piperazine-1-carboxylicacid benzyl ester

2-Methanesulfonyl-thieno[2,3-h]quinazoline-8-carboxylic acid (3.51 g,11.38 mmol), 1-hydroxyazatriazole (1.70 g, 12.52 mmol),1-benzylpiperazine carboxylate (2.76 g, 12.52 mmol) anddiisopropylethylamine (1.62 g, 12.52 mmol) were dissolved in dry DMF (35mL) and cooled in an ice-bath. EDC (2.40 g, 12.52 mmol) was added in oneportion and the resulting suspension was stirred at 0° C. for 20 minutesand for a further 16 hours at room temperature. The reaction wasconcentrated under reduced pressure and partitioned between hot DCM andbrine. The aqueous layer was extracted with DCM (3×50 mL) and thecombined organics were washed sequentially with dilute HCl (1×50 mL),saturated Na₂CO₃ (1×50 mL) and brine (1×50 mL), dried over Na₂SO₄,filtered and concentrated under reduced pressure. The yellow waxobtained was subjected to column chromatography (50% EtOAc in DCM,loaded in DCM, ˜350 mL silica) giving a cream solid which wasimmediately triturated with EtOAc, filtered and washed with pentane(3×20 mL) to give a cream powder (4.33 g, 75% yield). The filtrationliquors were concentrated under reduced pressure and the solid obtainedwas triturated with EtOAc, filtered and washed with pentane (3×5 mL) togive a second crop of cream powder (0.58 g, 10% yield). MS (ES⁺) 511. δH(CDCl₃) 3.5 (3H, s), 3.7 (4H, br m), 3.9 (4H, br m), 7.3-7.4 (5H, m),8.0 (1H, d), 8.2-8.3 (1H, d), 8.5 (1H, s), 9.6 (1H, s).

Example 16

4-[2-((R)-1-Phenyl-ethylamino)-thieno[2,3-h]quinazoline-8-carbonyl]-piperazine-1-carboxylicacid benzyl ester

4-(2-Methanesulfonyl-thieno[2,3-h]quinazoline-8-carbonyl)-piperazine-1-carboxylicacid benzyl ester (0.2 g, 0.39 mmol), TFA (4.5 mg, 0.04 mmol) and(R)-(α-methyl)benzylamine (0.47 g, 3.92 mmol) was suspended/dissolved indry dioxane (2 mL) and stirred at 120° C. for 16 hours. The reactionmixture was concentrated under reduced pressure and partitioned betweenDCM and dilute HCl. The organic layer was washed with saturated Na₂CO₃(1×10 mL) and brine (1×10 mL), dried over Na₂SO₄, filtered andconcentrated under reduced pressure. The brown gum obtained wasredissolved in DCM, silica (˜5 mL) was added and the suspension wasconcentrated under reduced pressure. The solid obtained was subjected tocolumn chromatography (25% EtOAc in DCM, ˜75 mL silica) to give a lightyellow gum (198.4 mg, 92% yield). MS (ES⁺) 552, (ES⁻) 551. δH (CDCl₃)1.7 (3H, d), 3.6-3.7 (4H, br m), 3.8-4.0 (4H, br m), 5.2 (2H, s), 5.4(1H, m), 5.7-5.8 (1H, br d), 7.2-7.5 (10H, m), 7.6 (2H, s), 8.1-8.2 (1H,s), 9.0 (1H, s)

Example 17

4-[2-(3-Piperidin-1-yl-phenylamino)-thieno[2,3-h]quinazoline-8-carbonyl]-piperazine-1-carboxylicacid benzyl ester

Formyl[(3-piperadinyl)-aniline] (0.14 g, 0.71 mmol) was dissolved in dryTHF (5 mL) and sodium hydride (35 mg, 0.88 mmol, 60 wt % in oil) wasadded in one portion. The resultant suspension was stirred at roomtemperature for 1 hour and cooled in an ice-bath.4-(2-Methanesulfonyl-thieno[2,3-h]quinazoline-8-carbonyl)-piperazine-1-carboxylicacid benzyl ester (0.3 g, 0.59 mmol) was added in one portion and thesuspension obtained was stirred at 0° C. for 1.5 hours. HCl (1 mL, 6M)was added and the homogeneous reaction was allowed to stir at roomtemperature for 3 hours. After concentration under reduced pressure thereaction mixture was partitioned between DCM and dilute HCl. The organiclayer was washed with very dilute Na₂CO₃ (High dilution reduced emulsionproblems), brine (1×10 mL), dried over Na₂SO₄, filtered and concentratedunder reduced pressure. The yellow gum obtained was subjected to columnchromatography (5% MeOH in DCM, loaded in DCM, ˜50 mL silica) and gave abright yellow powder (212 mg, 59% yield). MS (ES⁺) 607, (ES⁻) 605. δH(CDCl₃) 1.6 (2H, m), 1.8 (4H, m), 3.3 (4H m), 3.6-3.7 (4H, br m),3.8-4.0 (4H, br m), 5.2 (2H, s), 6.7 (1H, m), 7.3 (3H, m), 7.4 (6H, m),7.6 (1H, m), 7.7 (2H, m), 8.3 (1H, s), 9.1 (1H, s).

Example 18

Piperazin-1-yl-[2-(3-piperidin-1-yl-phenylamino)-thieno[2,3-h]quinazolin-8-yl]-methanone

4-[2-(3-Piperidin-1-yl-phenylamino)-thieno[2,3-h]quinazoline-8-carbonyl]-piperazine-1-carboxylicacid benzyl ester (205 mg, 0.34 mmol) was dissolved in dry DCM (4 mL)and HBr (2 mL, 33 wt % solution in AcOH) added. The resultant mixturewas stirred at room temperature for 0.5 hours and concentrated underreduced pressure. The yellow gum obtained was partitioned between Et₂Oand dilute HCl. The aqueous layer was extracted with Et₂O (1×10 mL). Theaqueous layer was then basified with saturated Na₂CO₃ and extracted withDCM (3×10 mL). The DCM organics were dried over Na₂SO₄, filtered andconcentrated under reduced pressure. The brown gum obtained wasredissolved in DCM, silica (˜5 mL) added and concentrated under reducedpressure. The solid obtained was subjected to column chromatography (10%MeOH in DCM, ˜100 mL silica) to give a bright yellow solid. This solidwas triturated with Et₂O, filtered and washed with Et₂O (3×5 mL) andpentane (3×5 mL) to give a yellow powder (96.4 mg, 60% yield). MS (ES⁺)473, (ES⁻) 472. δH (CDCl₃) 1.5-1.9 (6H, m), 2.9-3.1 (4H, m), 3.2-3.3(4H, m), 3.8-3.9 (4H, m), 6.7 (1H, m), 7.2-7.3 (3H, m), 7.4 (1H, s),7.6-7.7 (3H, m), 8.2-8.3 (1H, s), 9.1 (1H, s)

Example 19

4-Oxo-4,5,6,7-tetrahydro-benzo[b]thiophene-2-carboxylic acid tert-butylester

4-Oxo-4,5,6,7-tetrahydro-benzo[b]thiophene-2-carboxylic acid (Behringer,H.; Falkenberg, K. Chem. Ber., 1966, 99, 3309) (7.69 g, 39.19 mmol) wassuspended in 120 mL 2:1 cyclohexane: dichloromethane. Tert-butyl2,2,2-trichloroacetimidate (17.13 g, 78.38 mmol) was added in oneportion, followed by boron trifluoride diethyl etherate (87.8 mg, 6.19μmol, 0.16 mol %). The resultant slurry was stirred vigorously for 1.5hours and a further portion of tert-butyl 2,2,2-trichloroacetimidate(8.57 g, 39.19 mmol) was added. Stirring was continued overnight.Further portions of tert-butyl 2,2,2-trichloroacetimidate (8.57 g, 39.19mmol) were added at 1.5 hours and 3 hours and after a further 3 hoursthe reaction was quenched by cautious addition of solid NaHCO₃ (˜1 g).Silica was added (˜80 mL) and the resulting suspension was concentratedunder reduced pressure. The resulting crude mixture was purified bysilica gel chromatography (20% EtOAc in hexanes on 1 L of silica) toafford the title compound as a light yellow solid (8.81 g, 89% yield).MS (ES⁺) 253. δH (CDCl₃) 1.5-1.6 (9H, s), 2.2-2.3 (2H, m), 2.5-2.6 (2H,m), 3.0-3.1 (2H, m), 7.9 (1H, s).

Example 20

5-Dimethylaminomethylene-4-oxo-4,5,6,7-tetrahydro-benzo[b]thiophene-2-carboxylicacid tert-butyl ester

4-Oxo-4,5,6,7-tetrahydro-benzo[b]thiophene-2-carboxylic acid tert-butylester (8.75 g, 34.68 mmol) was suspended in dry toluene (40 mL) andBredereck's reagent (6.65 g, 38.14 mmol) was added. The resultingsuspension was heated at reflux for 2 hours, during which time allstarting material dissolved, allowed to cool to room temperature andconcentrated under reduced pressure. The resulting oil was purified bysilica gel chromatography (50% EtOAc in hexanes on 1 L silica, loaded asa solution in dichloromethane) to afford the title compound as a brownsolid (6.21 g, 58% yield). MS (ES⁺) 308. δH (CDCl₃) 1.5-1.6 (9H, s),2.9-3.0 (2H, m), 3.0-3.1 (2H, m), 3.1 (6H, s), 7.6 (1H, s), 8.0 (1H, s).

Example 21

2-(3-Sulfamoyl-phenylamino)-5,6-dihydro-thieno[2,3-h]quinazoline-8-carboxylicacid tert-butyl ester

5-Dimethylaminomethylene-4-oxo-4,5,6,7-tetrahydro-benzo[b]thiophene-2-carboxylicacid tert-butyl ester (6.21 g, 20.20 mmol),3-guanidinophenylsulphonamide hydrochloride (5.07 g, 20.20 mmol) andsodium hydroxide (0.81 g, 20.20 mmol) were suspended in isopropanol (250mL) and stirred at reflux overnight. The reaction was allowed to cool toroom temperature and diluted with water (˜200 mL). The resultingprecipitate was isolated by filtration and the solid obtained was washedwith water (1×100 mL), isopropanol (1×50 mL), diethyl ether (2×50 mL)and pentane (3×100 mL). Air drying gave an ochre powder (4.92 g, 53%yield). MS (ES⁺) 459. δH (d⁶ DMSO) 1.5-1.6 (9H, s), 2.9-3.0 (2H, m),3.1-3.2 (2H, m), 7.2-7.3 (2H, br s), 7.3-7.5 (2H, m), 7.7-7.8 (1H, m),8.1 (1H, s), 8.4 (1H, s), 8.6 (1H, s), 9.9 (1H, s).

Example 22

2-(3-Sulfamoyl-phenylamino)-thieno[2,3-h]quinazoline-8-carboxylic acidtert-butyl ester

2-(3-Sulfamoyl-phenylamino)-5,6-dihydro-thieno[2,3-h]quinazoline-8-carboxylicacid tert-butyl ester (4.90 g, 10.69 mmol) and2,3-dichloro-5,6-dicyano-1,4-benzoquinone (4.85 g, 21.37 mmol) weresuspended in dry 1,4-dioxane (300 mL) and heated at reflux overnight.After allowing the reaction to cool to room temperature the mixture wasconcentrated under reduced pressure and partitioned between ethylacetate and 1:1 saturated Na₂CO₃:brine. The organic layer was washedwith further portions of 1:1 saturated Na₂CO₃:brine (2×200 mL), driedover Na₂SO₄, and filtered. Silica (˜0.60 mL) was added to the filtrateand the resulting suspension was concentrated under reduced pressure.The resulting solid was purified by silica gel chromatography (80-90%EtOAc in hexanes on ˜800 mL of silica) to afford the title compound as ayellow solid (1.84 g, 38% yield) and further impure material (˜2 g). MS(ES⁺) 457. δH (d⁶ DMSO) 1.6 (9H, s), 7.3 (2H, br s), 7.4-7.6 (2H, m),7.8-7.9 (1H, m), 8.0 (2H, m), 8.7 (1H, s), 9.1 (1H, br s), 9.4 (1H, s),10.4-10.5 (1H, s).

A variety of other compounds of formula V have been prepared by methodssubstantially similar to those described in Example 22. Thecharacterization data for these compounds is summarized in Table 14below and includes HPLC, LC/MS (observed) and ¹H NMR data.

¹H NMR data is summarized in Table 14 below wherein ¹H NMR data wasobtained at 400 MHz in CDCl₃, unless otherwise indicated, and was foundto be consistent with structure. Compound numbers correspond to thecompound numbers listed in Table 5.

TABLE 14 Characterization Data for Selected Compounds of Formula V Com-M + 1 Rt pound No. (obs) (min) ¹H-NMR V-29 434 11.7 (CDCl₃) 1.5 (9H, s),1.6-1.7 (9H, s), 7.2 (1H, m), 7.3-7.4 (1H, m), 7.4 (1H, s), 7.5-7.6 (1H,m), 7.6-7.7 (1H, m), 8.0-8.1 (1H, s), 8.7-8.8 (1H, s), 9.1 (1H, s) V-30454 11.6 1.6-1.7 (9H, s), 7.4 (2H, m), 7.5 (4H, m), 7.6-7.7 (4H, m),7.8-7.9 (1H, m), 8.1-8.2 (1H, s), 8.7-8.8 (1H, s), 9.1 (1H, s) V-31 43411.6 1.4 (9H, s), 1.7 (9H, s), 7.4 (1H, m), 7.4-7.5 (2H, m), 7.7 (2H,s), 7.8 (2H, m), 8.7-8.8 (1H, s), 9.1 (1H, s) V-32 396 11.2 1.7 (9H, s),6.8 (1H, m), 7.3-7.4 (2H, m), 7.5 (1H, s), 7.7 (2H, m), 8.0-8.1 (1H, m),8.7-8.8 (1H, s), 9.1 (1H, s) V-33 392 11.1 1.7 (9H, s), 2.4-2.5 (3H, s),7.1 (1H, m), 7.2 (1H, s), 7.3 (1H, m), 7.3-7.4 (1H, m), 7.7 (2H, s),8.4-8.5 (1H, m), 8.7 (1H, s), 9.1 (1H, s) V-34 378 11.0 (DMSO-d6) 1.70(9H, s), 7.05 (1H, t), 7.40 (2H, t), 8.00 (1H, s), 8.05 (2H, d), 8.60(1H, d), 9.40 (1H, s), 10.15 (1H, s) V-35 / 9.5 1.10-1.90 (17H, m), 2.00(2H, m), 3.75 (1H, m), 6.95 (1H, d), 7.25 (1H, d), 8.05 (1H, s), 8.60(1H, d)

Example 23

2-(3-Sulfamoyl-phenylamino)-thieno[2,3-h]quinazoline-8-carboxylic acid

2-(3-Sulfamoyl-phenylamino)-thieno[2,3-h]quinazoline-8-carboxylic acidtert-butyl ester (1.84 g, 4.02 mmol) was placed in a 100 mL florentineand cooled in an ice-bath. Premixed TFA/dichloromethane/water (1:4:0.1,9 mL TFA) was added in one portion and the resultant suspension wasstirred at 0° C. for 10 minutes. After a further 2.5 hours at roomtemperature and 1.5 hours at 40° C. the reaction was concentrated underreduced pressure. The solid obtained was azeotroped with drydichloromethane (3×10 mL), triturated with diethyl ether and isolated byfiltration. The solid obtained was washed with diethyl ether (4×5 mL)and dried in a drying pistol at 40° C. overnight to afford the titlecompound as a yellow powder (1.44 g, 86% yield containing 1.24 eq. TFA).MS (ES⁺) 400. δH (d⁶ DMSO) 7.2-7.4 (2H, br s), 7.5 (1H, m), 7.6 (1H, m),8.0 (3H, m), 8.7 (1H, s), 8.8-8.9 (1H, s), 9.4 (1H, s), 10.4-10.5 (1H,s), 13.4-13.8 (1H, br s).

Example 24

(2-{[2-(3-Sulfamoyl-phenylamino)-thieno[2,3-h]quinazoline-8-carbonyl]-amino}-ethyl)-carbamicacid tert-butyl ester

2-(3-Sulfamoyl-phenylamino)-thieno[2,3-h]quinazoline-8-carboxylic acid(150 mg, 0.37 mmol), 1-hydroxy-7-azabenzotriazole (56 mg, 0.41 mmol),N,N-diisopropylethylamine (73 mg, 0.56 mmol) and BOC-ethylenediamine (66mg, 0.41 mmol) were dissolved in dry DMF (2 mL) and cooled in anice-bath. EDC (79 mg, 0.41 mmol) was added in one portion and theresultant suspension was stirred overnight allowing the ice-bath tomelt. The reaction was concentrated under reduced pressure, redissolvedin isopropanol (˜10 mL) and water was added (˜5 mL). The precipitateformed was isolated by filtration and washed with water (1×5 mL), diluteNaHSO₄ (1×5 mL), water (1×5 mL), saturated Na₂CO₃ (1×5 mL), water (1×5mL), isopropanol (1×5 mL), diethyl ether (2×5 mL) and pentane (2×5 mL)giving an ochre powder (102 mg, 51% yield). MS (ES⁺) 543. δH (d⁶ DMSO)1.2-1.5 (9H, s), 3.3-3.5 (4H, br m), 6.6 (0.1H, br s), 7.0 (0.9H, br s),7.1-7.4 (2H, br s), 7.4-7.6 (2H, m), 7.8-8.0 (2H, m), 8.2 (1H, m),8.6-9.0 (3H, br m), 9.4 (1H, s), 10.3-10.6 (1H, br s).

Example 25

2-(3-Sulfamoyl-phenylamino)-thieno[2,3-h]quinazoline-8-carboxylic acid(2-amino-ethyl)-amide

(2-{[2-(3-Sulfamoyl-phenylamino)-thieno[2,3-h]quinazoline-8-carbonyl]-amino}-ethyl)-carbamicacid tert-butyl ester (138.1 mg, 0.25 mmol) was placed in a 50 mLflorentine and cooled in an ice-bath. Premixed TFA/dichloromethane/water(1:4:0.1, 0.5 mL TFA) was added in one portion and the resultantsuspension was stirred at 0° C. for 35 minutes. After a further 1.75hours at room temperature the solution was concentrated under reducedpressure to give a yellow solid. This solid was azeotroped withdichloromethane (3×10 mL), triturated with diethyl ether and isolated byfiltration. The solid collected was washed with diethyl ether (3×5 mL)and pentane (3×5 mL) and placed in a drying pistol at 40° C. overnightgiving a yellow powder (145.5 mg, 95% yield containing ˜1.5 eq. TFA). MS(ES⁺) 442. δH (d⁶ DMSO) 3.0-3.1 (2H, m), 3.5-3.7 (2H, m), 7.3-7.4 (2H,br s), 7.5 (1H, m), 7.6 m), 7.8-8.1 (5H, m), 8.2-8.3 (1H, m), 8.7-8.9(3H, m), 9.4 (1H, s), 10.5 (1H, s).

A variety of other compounds of formula V have been prepared by methodssubstantially similar to those described in Example 18 or 25. Thecharacterization data for these compounds is summarized in Table 15below and includes HPLC, LC/MS (observed) and ¹H NMR data.

¹H NMR data is summarized in Table 15 below wherein ¹H NMR data wasobtained at 400 MHz in deuterated DMSO, unless otherwise indicated, andwas found to be consistent with structure. Compound numbers correspondto the compound numbers listed in Table 5.

TABLE 15 Characterization Data for Selected Compounds of Formula V Com-pound M + 1 Rt No. (obs) (min) ¹H-NMR V-36 468.0 8.5 1.5-1.8 (6H, br m),3.5-3.9 (4H, br m), 7.1-7.4 (2H, br s), 7.4-7.6 (2H, m), 7.8-8.0 (3H,m), 8.3 (1H, s), 9.2 (1H, s), 9.4 (1H, s), 10.3-10.6 (1H, br s) V-37470.0 7.5 3.5-4.0 (8H, br m), 7.2-7.4 (2H, br s), 7.4-7.5 (2H, m), 7.7(1H, m), 7.9 (1H, m), 8.0 (1H, m), 8.4 (1H, s), 9.3-9.4 (2H, m),10.3-10.6 (1H, br s) V-38 569.0 8.9 1.4-1.5 (9H, s), 3.4-3.6 (4H, br m),3.6-4.0 (4H, br m), 7.3 (2H, s), 7.4-7.6 (2H, m), 7.7-7.8 (1H, m), 7.9(1H, m), 8.0 (1H, m), 8.4 (1H, s), 9.3 (1H, s), 9.4 (1H, s), 10.4-10.5(1H, s) V-39 557.0 8.7 1.2-1.5 (9H, s), 1.6-1.8 (2H, m), 3.0-3.1 (2H,m), 3.2-3.5 (2H, m), 6.5 (0.1H, br s), 6.8-6.9 (0.9H, br s), 7.1-7.4(2H, br s), 7.4-7.6 (2H, m), 7.8-8.0 (2H, m), 8.0-8.2 (1H, m), 8.6-8.8(1H, br s), 8.8 (1H, s), 9.0 (1H, br s), 9.3-9.5 (1H, br s), 10.3-10.6(1H, br s) V-40 483.0 7.5 2.6-2.8 (3H, br m), 2.9-4.8 (8H, br m), 7.4(2H, br s), 7.5 (2H, m), 7.7-7.8 (1H, m), 7.9-8.1 (2H, m), 8.4 (1H, s),9.4 (2H, m), 10.5 (1H, s) V-41 557.0 8.8 1.3 (9H, s), 2.8-2.9 (3H, m),3.3-3.5 (4H, m), 7.2-7.4 (2H, br s), 7.5-7.6 (2H, m), 7.8-8.0 (2H, m),8.1-8.3 (1H, m), 8.7-8.9 (3H, m), 9.4 (1H, s), 10.4 (1H, s) V-42 471.07.3 2.1-2.3 (6H, m), 3.2-3.6 (4H, m), 7.0-7.3 (2H, br s), 7.4-7.6 (2H,m), 7.8-8.1 (3H, m), 8.5-8.7 (1H, br s), 8.8-8.9 (1H, s), 9.1 (1H, s),9.4 (1H, s), 10.2-10.6 (1H, br s) V-4 468.9 6.8 2.7-2.9 (4H, br m),3.5-4.0 (4H, br m), 7.2-7.4 (2H, br s), 7.4-7.6 (2H, m), 7.7-7.8 (1H,m), 7.8-8.0 (2H, m), 8.3-8.4 (1H, br s), 9.2-9.3 (1H, br s), 9.3-9.5(1H, br s), 10.3-10.5 (1H, s) V-43 457.0 6.8 2.3-2.4 (3H, s), 2.7-2.8(2H, m), 3.4-3.5 (2H, m), 5.4-6.2 (2H, br s), 7.5 (1H, m), 7.5-7.6 (1H,m), 7.9 (1H, m), 8.0 (1H, m), 8.1 (1H, m), 8.5 (1H, m), 8.7-8.8 (1H, s),9.0-9.1 (1H, s), 9.4-9.5 (1H, s), 10.5 (1H, s) V-44 444.0 6.6 3.0-3.1(2H, m), 3.5-3.7 (2H, m), 7.3-7.4 (2H, br s), 7.5 (1H, m), 7.6 (1H, m),7.8-8.1 (5H, m), 8.2-8.3 (1H, m), 8.7-8.9 (3H, m), 9.4 (1H, s), 10.5(1H, s) V-45= 458.0 6.8 1.8-2.0 (2H, m), 2.8-3.0 (2H, m), 3.3-3.5 (2H,m), 7.3-7.4 (2H, br s), 7.5 (1H, m), 7.5-7.6 (1H, m), 7.6-7.8 (3H, brs), 7.8-8.0 (2H, m), 8.1-8.2 (1H, m), 8.7-8.8 (2H, m), 8.9 (1H, s), 9.4(1H, s), 10.4-1.5 (1H, s) V-46 583.0 8.7 1.2-1.5 (9H, s), 1.8-2.0 (2H,m), 2.8-3.7 (5H, m), 4.1-4.5 (2H, m), 6.5-7.0 (1H, m), 7.1-7.4 (2H, brs), 7.4-7.6 (2H, m), 7.8-8.0 (3H, m), 8.3-8.4 (1H, s), 9.2 (1H, s), 9.4(1H, s), 10.4-10.5 (1H, br s) V-47 568.9 8.5 1.2-1.5 (9H, m), 1.8-2.3(2H, m), 3.4-3.9 (2H, m), 4.0-4.3 (3H, m), 6.8-7.4 (3H, br m), 7.4-7.6(2H, m), 7.7-8.0 (3H, m), 8.4-8.6 (1H, m), 9.0-9.3 (1H, m), 9.4 (1H, s),10.3-10.6 (1H, br s) V-48 484.0 6.4 1.4-1.6 (2H, m), 1.9-2.1 (2H, m),2.8-3.8 (3H, m), 4.2-4.7 (2H, m), 7.2-7.4 (2H, br s), 7.4-7.6 (2H, m),7.7-8.1 (6H, m), 8.4 (1H, s), 9.2 (1H, s), 9.4 (1H, s), 10.4-10.5 (1H,br s) V-49 470.0 6.6 2.0-2.5 (3H, m), 3.6-4.6 (4H, m), 7.3-7.9 (5H, m),7.9-8.3 (5H, m), 8.4-8.6 (1H, m), 9.2-9.3 (1H, m), 9.4 (1H, s), 10.5(1H, br s) V-50 404 9.0 2.25 (3H, s), 2.40 (2H, t), 3.80 (2H, m), 7.05(1H, t), 7.40 (2H, t), 7.85-8.15 (4H, m), 8.25 (1H, s), 9.40 (1H, s),10.10 (1H, s) V-51 406 7.9 3.69 (2H, m), 3.76 (2H, m), 5.12 (2H, s),6.23 (1H, m), 6.96 (2H, m), 7.52 (1H, s), 7.84 (1H, d), 7.90 (1H, d),8.36 (1H, s), 9.31 (1H, s) and 9.80 (1H, s) V-52 391 9.0 3.65-3.90 (8H,2 × m), 7.05 (1H, t), 7.40 (2H, t), 7.80-8.10 (4H, dd and d), 8.30 (1H,s), 9.40 (1H, s), 10.10 (1H, s) V-53 490 9.8 1.30-1.50 (9H, m),1.80-2.25 (2H, m), 3.40-3.80 (3H, m), 4.10-4.30 (2H, m), 7.05 (1H, q),7.40 (2H, m), 7.85-8.15 (4H, dd and d), 8.40-8.55 (1H, m), 9.40 (1H, s),10.15 (1H, s) V-54 390 8.2 2.00-2.80 (2H, m), 3.60-4.50 (5H, m), 7.05(1H, q), 7.45 (2H, m), 7.90-8.30 (6H, m), 8.40-8.55 (1H, m), 9.40 (1H,s), 10.15 (1H, s) V-55 447 10.2 1.3-1.4 (9H, s), 3.6-3.8 (8H, br m),7.0-7.1 (1H, m), 7.3 (1H, m), 7.7-7.8 (1H, m), 7.8-7.9 (1H, m), 7.9-8.0(1H, m), 8.1-8.3 (2H, m), 9.3-9.4 (1H, s), 10.0 (1H, s) V-56 460 10.31.3 (9H, s), 2.2 (3H, s), 2.3-2.4 (4H, m), 3.6-3.8 (4H, m), 7.0-7.1 (1H,m), 7.3 (1H, m), 7.7 (1H, m), 7.8-7.9 (1H, m), 7.9-8.0 (1H, m), 8.2-8.3(2H, m), 9.3-9.4 (1H, s), 10.0 (1H, s) V-57 546 10.6 1.3-1.5 (18H, m),1.8-2.3 (2H, m), 3.4-4.3 (5H, m), 7.0-7.1 (1H, m), 7.2-7.4 (2H, m),7.6-7.8 (1H, m), 7.8-8.0 (2H, m), 8.0-8.2 (1H, m), 8.3-8.5 (1H, m),9.3-9.4 (1H, s), 10.0 (1H, s) V-58 467 10.0 3.4-3.8 (8H, br m), 7.3 (1H,m), 7.4-7.6 (4H, m), 7.7 (2H, m), 7.8-8.0 (3H, m), 8.2 (1H, s), 8.6 (1H,br s), 9.4 (1H, s), 10.2 (1H, s) V-59 480 10.1 2.2 (3H, s), 2.2-2.4 (4H,br s), 3.6-3.7 (4H, br s), 7.3 (1H, m), 7.3-7.6 (4H, m), 7.7 (2H, m),7.8-8.0 (3H, m), 8.1-8.2 (1H, s), 8.6-8.7 (1H, br s), 9.4 (1H, s), 10.2(1H, s) V-60 566 10.5 1.2-1.5 (9H, m), 1.7-2.1 (2H, m), 3.3-4.2 (5H, m),7.2-7.3 (2H, m), 7.4-7.5 (4H, m), 7.7 (2H, m), 7.8-8.0 (3H, m), 8.3-8.4(1H, m), 8.4-8.6 (1H, m), 9.4 (1H, s), 10.2-10.3 (1H, m) V-61 409 9.23.6-3.9 (8H, br m), 6.8-6.9 (1H, m), 7.4 (1H, m), 7.7-7.8 (1H, m), 7.9(1H, m), 8.0 (1H, m), 8.1 (1H, m), 8.2 (1H, s), 9.4 (1H, s), 10.3-10.4(1H, s) V-62 422 9.4 2.2-2.3 (3H, br s), 2.4 (4H, m), 3.7-3.8 (4H, brs), 6.8 (1H, m), 7.3-7.4 (1H, m), 7.7 (1H, m), 7.9 (1H, m), 8.0 (1H, m),8.2 (2H, m), 9.4 (1H, s), 10.3-10.4 (1H, s) V-63 508 9.9 1.3-1.5 (9H, brm), 1.8-2.3 (2H, m), 3.4-4.2 (5H, m), 6.8 (1H, m), 7.3-7.5 (2H, m),7.7-7.9 (1H, m), 7.9 (1H, m), 8.0 (1H, m), 8.0-8.2 (1H, m), 8.4-8.5 (1H,m), 9.4 (1H, s), 10.3-10.4 (1H, s) V-64 405 9.1 2.3 (3H, s), 3.6-3.8(8H, br m), 7.1 (1H, m), 7.2-7.3 (2H, m), 7.7-7.8 (1H, m), 7.8-7.9 (2H,m), 8.0-8.1 (1H, s), 9.2 (1H, s), 9.3 (1H, s) V-65 418 9.2 2.2-2.3 (3H,br s), 2.3 (3H, br s), 2.3-2.4 (4H, br m), 3.5-3.8 (4H, br m), 7.1 (1H,m), 7.2-7.3 (2H, m), 7.7-7.8 (1H, m), 7.8-7.9 (2H, m), 8.0-8.1 (1H, s),9.2 (1H, s), 9.3 (1H, s) V-66 504 9.9 1.3-1.5 (9H, br m), 1.8-2.3 (2H,m), 2.3-2.4 (3H, s), 3.4-4.3 (5H, m), 7.1 (1H, m), 7.2-7.4 (3H, m),7.7-7.8 (1H, m), 7.8-7.9 (2H, m), 8.2-8.4 (1H, m), 9.2-9.3 (2H, m) V-67545 8.4 1.39 (2H, m), 1.50 (4H, m), 2.37 (4H, m), 2.53 (2H, m), 2.63(2H, m), 3.69 (4H, m), 3.80 (4H, m), 7.00 (1H, d), 7.23 (1H, t), 7.33(1H, d), 7.89 (1H, d), 7.98 (1H, d), 8.73 (1H, s), 9.07 (1H, br s), 9.37(1H, s), 10.07 (1H, s) and 10.29 (1H, s) V-68 448 8.0 2.10 (3H, s), 3.69(4H, m), 3.81 (4H, m), 7.00 (1H, d), 7.20 (1H, t), 7.28 (1H, d), 7.85(1H, d), 7.93 (1H, d), 8.73 (1H, s), 9.10 (1H, br s), 9.25 (1H, s),10.04 (1H, s) and 10.07 (1H, s) V-69 547 8.1 2.37 (4H, m), 2.50 (2H, m),2.66 (2H, m), 3.56 (4H, m), 3.65 (4H, m), 3.80 (4H, m), 7.03 (1H, d),7.23 (1H, t), 7.30 (1H, d), 7.88 (1H, d), 7.95 (1H, d), 8.71 (1H, s),9.03 (1H, br s), 9.37 (1H, s), 10.08 (1H, s) and 10.14 (1H, s) V-70 5608.0 2.12 (3H, s), 2.20-2.60 (10H, br m), 2.69 (2H, m), 3.67 (4H, m),3.79 (4H, m), 7.02 (1H, d), 7.23 (1H, t), 7.32 (1H, d), 7.88 (1H, d),7.96 (1H, d), 8.73 (1H, s), 9.09 (1H, br s), 9.35 (1H, s), 10.07 (1H, s)and 10.18 (1H, s) V-71 505 8.0 2.84 (6H, s), 2.89 (2H, m), 3.45 (2H, m),3.70 (4H, m), 3.77 (4H, m), 7.21 (1H, d), 7.28 (1H, t), 7.47 (1H, d),7.89 (1H, d), 7.97 (1H, d), 8.63 (1H, s), 8.77 (1H, br s), 9.37 (1H, s),9.40 (1H, br s), 10.13 (1H, s) and 10.29 (1H, s) V-72 446 9.8 1.3-1.4(9H, s), 2.0-2.7 (2H, m), 3.6-4.3 (5H, m), 7.0-7.1 (1H, m), 7.3 (1H, m),7.7-8.2 (6H, m), 8.3-8.5 (1H, m), 9.3-9.4 (1H, s), 10.0 (1H, s) V-73 4669.6 2.0-2.4 (2H, m), 3.5-4.1 (5H, m), 7.3 (1H, m), 7.4-7.6 (4H, m), 7.7(1H, m), 7.8-8.2 (6H, m), 8.3-8.5 (2H, m), 9.4 (1H, s), 10.2-10.3 (1H,m) V-74 408 8.6 2.0-2.6 (2H, m), 3.6-4.4 (5H, m), 6.8 (1H, m), 7.4 (1H,m), 7.6-8.3 (6H, m), 8.3-8.5 (1H, m), 9.4 (1H, s), 10.4 (1H, s) V-75 4048.4 2.0-2.4 (2H, m), 2.3 (3H, s), 3.6-4.2 (5H, m), 7.1 (1H, m), 7.2-7.3(2H, m), 7.7-8.3 (7H, m), 9.2-9.4 (2H, m) V-76 535 7.8 2.83 (2H, s),3.20 (2H, m), 3.29 (5H, m), 3.60 (2H, m), 3.69 (4H, m), 3.77 (4H, m),7.19 (1H, d), 7.29 (1H, t), 7.50 (1H, d), 7.89 (1H, d), 7.97 (1H, d),8.55 (2H, br s), 8.62 (1H, s), 8.74 (1H, s), 9.36 (1H, s), 10.11 (1H, s)and 10.23 (1H, s) V-77 549 7.9 1.80 (2H, m), 2.80 (2H, m), 3.03 (2H, m),3.23 (3H, s), 3.29 (2H, m), 3.39 (2H, m), 3.69 (4H, m), 3.77 (4H, m),7.20 (1H, d), 7.30 (1H, t), 7.49 (1H, d), 7.89 (1H, d), 7.97 (1H, d),8.40 (2H, br s), 8.62 (1H, s), 8.78 (1H, s), 9.36 (1H, s), 10.11 (1H, s)and 10.25 (1H, s) V-78 573 9.5 0.70-0.92 (10H, br m), 1.70 (2H, m), 1.83(2H, m), 3.41 (4H, m), 3.67 (4H, m), 3.79 (4H, m), 7.19 (1H, d), 7.30(1H, t), 7.47 (1H, d), 7.89 (1H, d), 7.97 (1H, d), 8.63 (1H, s), 8.78(1H, s), 9.27 (1H, br s), 9.36 (1H, s), 10.11 (1H, s) and 10.25 (1H, s)V-79 446 9.8 1.4 (9H, s), 3.2 (4H, br s), 3.9-4.0 (4H, br s), 7.0-7.1(1H, m), 7.3 (1H, m), 7.8-8.0 (3H, m), 8.1 (1H, s), 8.3 (1H, s), 8.8-9.0(2H, br s), 9.3-9.4 (1H, s), 10.0 (1H, s) V-80 466 9.7 3.1-3.3 (4H, brs), 3.8-4.0 (4H, br m), 7.3-7.5 (5H, m), 7.7 (2H, m), 7.9-8.0 (2H, m),8.1-8.2 (1H, m), 8.2-8.4 (2H, m), 8.8-9.0 (2H, br s), 9.4 (1H, s), 10.2(1H, s) V-81 432 9.6 1.30 (6H, d), 2.95 (1H, m), 3.30 (4H, br s), 4.00(4H, br s), 6.95 (1H, d), 7.35 (1H, t), 7.85-8.05 (4H, m), 8.30 (1H, s),8.90 (2H, br s), 9.40 (1H, s), 10.10 (1H, s) V-82 432 9.4 1.30 (6H, d),2.00-2.50 (2H, m), 2.95 (1H, m), 3.60-4.40 (5H, m), 6.95 (1H, d), 7.35(1H, m), 7.70-8.60 (8H, m), 9.40 (1H, s), 10.05 (1H, s) V-83 496 10.02.15 (3H, s), 2.40 (4H, br s), 3.75 (4H, br s), 6.65 (1H, d), 7.15 (3H,m), 7.45 (3H, m), 7.70 (1H, d), 7.85-8.10 (4H, m), 9.45 (1H, s), 10.25(1H, s) V-84 482 9.6 3.25 (4H, br s), 4.00 (4H, br s), 6.70 (1H, d),7.15 (3H, m), 7.45 (3H, m), 7.80-8.10 (4H, m), 8.95 (2H, br s), 9.45(1H, s), 10.25 (1H, s) V-85 482 9.5 2.00-2.50 (2H, m), 3.65-4.30 (5H,m), 6.65 (1H, m), 7.15 (2H, m), 7.45 (2H, m), 7.75-8.40 (6H, m), 9.40(1H, s), 10.30 (1H, s) V-86 494 10.2 2.45 (4H, br s), 3.35 (3H, s), 3.75(4H, br s), 4.05 (2H, s), 6.90 (1H, d), 7.10-7.50 (6H, m), 7.85-8.25(5H, m), 9.35 (1H, s), 10.10 (1H, s) V-87 480 9.8 3.25 (4H, br s), 3.95(4H, br s), 4.00 (2H, s), 6.90 (1H, d), 7.00-7.50 (6H, m), 7.80-8.30(5H, m), 8.95 (1H, s), 9.35 (1H, s), 10.10 (1H, s) V-88 480 9.71.90-2.40 (2H, m), 3.65-4.35 (7H, m), 6.85 (1H, m), 7.15-7.40 (5H, m),7.75-8.50 (8H, m), 9.35 (1H, s), 10.15 (1H, s) V-89 470 9.4 3.40 (4H, brs), 4.05 (4H, br s), 7.20 (1H, m), 7.35 (1H, t), 7.90-8.10 (3H, m), 8.30(1H, s), 8.55 (1H, s), 8.95 (2H, br s), 9.40 (1H, s), 10.35 (1H, s) V-90478 10.1 2.30 (3H, s), 3.35 (4H, br s), 3.95 (4H, br s), 7.05 (1H, d),7.20-7.55 (4H, m), 7.90-8.10 (3H, m), 8.25-8.40 (2H, m), 8.95 (2H, brs), 9.45 (1H, s), 10.25 (1H, s) V-91 494 9.6 3.30 (4H, br s), 3.85 (3H,s), 3.95 (4H, br s), 7.05 (1H, t), 7.20 (2H, m), 7.30-7.55 (3H, m),7.90-8.30 (4H, m), 8.95 (2H, br s), 9.45 (1H, s), 10.20 (1H, s) V-92 49410.4 2.10 (6H, s), 3.30 (4H, br s), 3.95 (4H, br s), 6.85 (1H, d), 7.20(3H, m), 7.55 (1H, t), 7.90-8.10 (2H, dd), 8.20 (1H, s), 8.35 (1H, d),9.00 (2H, br s), 9.45 (1H, s), 10.25 (1H, s) V-93 468 9.4 3.35 (4H, brs), 4.10 (4H, br s), 7.20 (1H, d), 7.35 (1H, t), 7.90-8.10 (4H, m), 8.35(1H, s), 8.60 (1H, s), 8.95 (2H, br s), 9.45 (1H, s), 10.35 (1H, s) V-94467 9.0 3.25 (4H, br s), 4.00 (4H, br s), 7.40 (1H, d), 7.60 (1H, t),7.70 (1H, m), 7.90-8.10 (2H, m), 8.20-8.50 (4H, m), 8.70 (1H, d),8.85-9.10 (3H, m), 9.45 (1H, s), 10.35 (1H, s) V-95 486 10.0 2.30 (3H,s), 3.25 (4H, br s), 4.05 (4H, br s), 6.95-7.25 (2H, m), 7.55 (2H, m),7.90-8.15 (3H, m), 8.30-8.45 (2H, m), 8.90 (2H, br s), 9.40 (1H, s),10.30 (1H, s) V-96 486 9.9 2.35 (3H, s), 3.25 (4H, br s), 4.05 (4H, brs), 6.95-7.40 (4H, m), 7.45 (2H, m), 7.90-8.10 (3H, m), 8.90 (2H, br s),9.40 (1H, s), 10.30 (1H, s) V-97 506 10.1 3.25 (4H, br s), 4.00 (4H, brs), 7.25-7.40 (2H, m), 7.50-7.70 (2H, m), 7.85-8.10 (2H, m), 8.20-8.40(3H, m), 8.90 (2H, br s), 9.45 (1H, s), 10.30 (1H, s) V-98 475 8.02.7-2.9 (4H, br m), 3.0-3.1 (4H, br m), 3.6-3.8 (8H, br m), 7.0 (2H, m),7.7-7.9 (4H, m), 8.1-8.2 (1H, s), 9.2-9.3 (1H, s), 9.8-9.9 (1H, s) V-99404 8.5 2.7-2.8 (4H, br m), 3.5-3.7 (4H, br m), 4.6-4.7 (2H, m), 7.1-7.2(1H, m), 7.2-7.5 (4H, m), 7.7-7.8 (2H, m), 8.0-8.3 (2H, m), 9.1-9.2 (1H,s) V-100 468 7.6 3.27 (4H, br s), 3.32 (3H, s), 4.03 (4H, br s), 7.58(1H, d), 7.65 (1H, t), 7.89 (1H, d), 7.97 (1H, d), 8.05 (1H, d), 8.45(1H, s), 8.90 (2H, br s), 9.46 (1H, s), 9.52 (1H, br s), 10.62 (1H, s)V-101 468 7.5 2.00-2.50 (2H, m), 3.28-3.32 (3H, m), 3.65-4.43 (5H, m),7.58-7.67 (2H, m), 7.82-8.10 (6H, m), 8.50-8.58 (1H, m), 9.34 (1H, brs), 9.45 (1H, s), 10.60 (1H, s) V-102 448 9.3 1.33 (6H, d), 3.27 (4H, brs), 3.95 (4H, br s), 4.63 (1H, quint.), 6.59 (1H, d), 7.28 (1H, t), 7.54(1H, s), 7.80 (1H, d), 7.92 (1H, d), 7.98 (1H, d), 8.29 (1H, s), 8.88(2H, br s), 9.38 (1H, s), 10.06 (1H, s) V-103 448 9.2 1.33 (6H, d),2.00-2.50 (2H, m), 3.60-4.25 (5H, m), 4.63 (1H, quint.), 6.60 (1H, m),7.20-7.33 (1H, m), 7.54-7.78 (2H, m), 7.90-8.07 (5H, m), 8.39-8.48 (1H,m), 9.38 (1H, s), 10.08 (1H, s) V-104 405 10.4 1.36 (9H, s), 3.34 (6H,s), 7.07 (1H, d), 7.30 (1H, t), 7.71 (1H, m), 7.89 (1H, d), 7.94 (1H,d), 8.23 (1H, br s), 8.31 (1H, s), 9.35 (1H, s), 9.99 (1H, s) V-105 4889.8 1.35 (9H, s), 2.05 (3H, s), 3.58 (4H, t), 3.70-3.82 (4H, m), 7.07(1H, d), 7.31 (1H, t), 7.82 (1H, m), 7.90 (1H, d), 7.96 (1H, d), 8.14(1H, t), 8.27 (1H, s), 9.36 (1H, s), 10.00 (1H, s) V-106 502 9.8 1.34(9H, s), 1.75-2.10 (5H, m), 3.55-3.95 (8H, m), 7.05-7.10 (1H, m), 7.32(1H, m), 7.86-8.10 (4H, m), 8.27 (1H, d), 9.35 (1H, s), 9.97-10.00 (1H,m) V-107 474 10.1 1.30-1.40 (11H, m), 1.83-1.89 (3H, m), 2.80-2.95 (2H,m), 3.20-3.45 (4H, m), 7.10 (1H, d), 7.34 (1H, t), 7.85-8.20 (5H, m),8.44 (1H, m), 8.70 (1H, s), 9.10 (1H, t), 9.34 (1H, s), 9.95 (1H, s)V-108 460 10.2 1.35 (9H, m), 1.71 (1H, m), 2.08 (1H, m), 2.40-2.62 (1H,m), 2.92-3.50 (6H, m), 7.11 (1H, d), 7.34 (1H, t), 7.86-8.01 (4H, m),8.55-8.70 (3H, m), 9.17 (1H, t), 9.35 (1H, s), 9.96 (1H, s) V-109 46010.3 1.34 (9H, m), 2.00-2.20 (2H, m), 3.20-4.20 (8H, m), 7.09 (1H, d),7.33 (1H, t), 7.88-8.04 (4H, m), 8.32 (1H, br s), 8.75 (2H, s), 9.36(1H, s), 10.00 (1H, s) V-110 446 9.9 1.35 (9H, s), 2.00-2.50 (2H, m),3.40-4.30 (5H, m), 7.08 (1H, d), 7.28-7.36 (1H, m), 7.77-8.10 (7H, m),8.40-8.48 (1H, m), 9.36 (1H, s), 10.02 (1H, s) V-111 446 9.9 1.35 (9H,s), 2.00-2.50 (2H, m), 3.30-4.30 (5H, m), 7.08 (1H, d), 7.28-7.36 (1H,m), 7.76-8.10 (7H, m), 8.40-8.47 (1H, m), 9.36 (1H, s), 10.02 (1H, s)V-112 475 8.4 (CDCl₃) 3.0 (4H, m), 3.2-3.3 (4H, m), 3.8-4.0 (8H, m), 6.7(1H, m), 7.2-7.5 (4H, m), 7.6 (1H, s), 7.7-7.8 (2H, m), 8.3 (1H, s),9.1-9.2 (1H, s) V-113 474 / 2.8 (4H, m), 3.0 (4H, m), 3.2 (4H, m), 3.7(4H, m), 6.4 (1H, m), 7.1-7.2 (1H, m), 7.3-7.4 (1H, m), 7.8-8.0 (3H, m),8.1-8.2 (1H, s), 9.3-9.4 (1H, s), 9.9-10.0 (1H, s) V-114 488 / 2.2 (3H,s), 2.8 (4H, m), 3.0-3.2 (4H, m), 3.2-3.6 (4H, br m), 3.6-3.8 (4H, m),7.0 (1H, m), 7.8-7.9 (4H, m), 8.2 (1H, s), 9.2-9.3 (1H, s), 9.8-9.9 (1H,s) V-115 474 / 2.8 (4H, m), 2.9 (4H, m), 3.1 (4H, m), 3.7 (4H, m),6.9-7.0 (2H, m), 7.8-7.9 (4H, m), 8.2 (1H, s), 9.2-9.3 (1H, s), 9.8-9.9(1H, s) V-116 391 7.8 2.9 (4H, m), 3.8-3.9 (4H, m), 7.9-8.1 (4H, m), 8.3(1H, m), 8.4-8.5 (2H, m), 9.4-9.5 (1H, s), 10.5 (1H, s) V-117 391 7.52.8 (4H, m), 3.6-3.8 (4H, m), 7.4-7.5 (1H, m), 7.9-8.1 (2H, m), 8.2 (2H,m), 8.5-8.6 (1H, m), 9.0-9.1 (1H, s), 9.4 (1H, s), 10.2-10.3 (1H, s)V-118 516 10.0 1.00-1.20 (2H, m), 1.35 (9H, s), 1.70-1.85 (3H, m), 1.99(3H, s), 2.40-2.55 (1H, m), 2.97-3.04 (1H, m), 3.10-3.30 (2H, m),3.80-3.84 (1H, m), 4.36-4.40 (1H, s), 7.10 (1H, d), 7.33 (1H, t),7.85-7.94 (3H, m), 8.00 (1H, t), 8.69 (1H, s), 9.04 (1H, t), 9.34 (1H,s), 9.94 (1H, s) V-119 502 9.9 1.34 (9H, s), 1.63-1.77 (1H, m),1.93-2.05 (4H, m), 2.43-2.55 (1H, m), 3.04-3.57 (6H, m), 7.10 (1H, d),7.34 (1H, dt), 7.86-7.98 (4H, m), 8.69 (1H, s), 9.10-9.13 (1H, m), 9.34(1H, s), 9.94 (1H, s) V-120 503 7.4 3.20-4.05 (16H, m) 7.50 (2H, d),7.85-8.45 (5H, m), 8.95 (2H, br s), 9.45 (1H, s), 10.40 (1H, s) V-121461 7.5 3.05 (6H, s), 3.25 (4H, br s), 4.00 (4H, br s), 7.50 (2H, d),7.90-8.45 (5H, m), 8.95 (2H, br s), 9.45 (1H, s), 10.40 (1H, s) V-122516 7.8 2.90 (3H, s), 3.00-4.00 (16H, m) 7.55 (2H, d), 7.90-8.40 (5H,m), 9.00 (2H, br s), 9.45 (1H, s), 10.45 (1H, s) V-123 418 8.8 (CDCl₃)1.6-1.7 (3H, d), 3.0 (4H, m), 3.8-3.9 (4H, m), 5.3-5.4 (1H, m), 5.7-5.8(1H, m), 7.2-7.4 (3H, m), 7.4-7.6 (4H, m), 8.1-8.2 (1H, s), 9.0 (1H, s)V-124 418 8.8 (CDCl₃) 1.6-1.7 (3H, d), 3.0 (4H, m), 3.8-3.9 (4H, m),5.3-5.4 (1H, m), 5.7-5.8 (1H, m), 7.2-7.4 (3H, m), 7.4-7.6 (4H, m),8.1-8.2 (1H, s), 9.0 (1H, s) V-125 418 9.0 (CDCl₃) 2.9-3.2 (6H, m),3.8-4.0 (6H, m), 5.4-5.5 (1H, m), 7.2-7.4 (5H, m), 7.6 (2H, m), 8.2-8.3(1H, s), 9.0 (1H, s) V-126 430 9.4 (CDCl₃) 1.9-2.1 (1H, m), 2.7-2.8 (1H,m), 2.9-3.2 (6H, m), 3.7-3.9 (4H, m), 5.5-5.7 (1H, m), 5.8-6.0 (1H, m),7.2-7.5 (4H, m), 7.6 (2H, m), 8.1-8.3 (1H, br s), 8.9-9.2 (1H, br s)V-127 430 9.0 (CDCl₃) 1.3-1.5 (2H, m), 2.1-2.2 (1H, m), 2.6-3.1 (4H, brm), 3.1-3.2 (1H, m), 3.5-3.9 (4H, br m), 5.7 (1H, s), 7.2-7.4 (5H, m),7.6 (2H, m), 8.0 (1H, s), 9.0 (1H, s) V-128 475 / 3.29 (3H, s), 7.15(1H, t), 7.41 (2H, t), 7.59 (1H, d), 7.70 (1H, t), 7.83 (2H, d), 7.96(1H, d), 8.04 (1H, d), 8.34 (1H, d), 8.95 (1H, s), 9.00 (1H, s), 9.44(1H, s), 10.50 (1H, s), 10.54 (1H, s) V-129 489 / 3.18 (3H, s), 4.55(2H, d), 7.27 (1H, t), 7.32-7.47 (4H, m), 7.56 (1H, d), 7.66 (1H, t),7.92 (1H, d), 8.00 (1H, d), 8.32 (1H, d), 8.82 (1H, s), 8.88 (1H, s),9.21 (1H, t), 9.41 (1H, s), 10.49 (1H, s) V-130 453 / 1.83-1.95 (2H, m),1.96-2.11 (2H, m), 3.26 (3H, s), 3.57 (2H, t), 4.02 (2H, t), 7.53-7.67(2H, m), 7.84 (1H, d), 7.90 (1H, d), 7.97 (1H, d), 8.51 (1H, s), 9.32(1H, s), 9.40 (1H, s), 10.53 (1H, s) V-131 482 / 3.26 (3H, s), 3.36 (2H,s), 3.97 (2H, s), 4.31 (2H, s), 7.56 (1H, d), 7.63 (1H, t), 7.88-7.98(2H, m), 8.02 (1H, d), 8.25 (1H, s), 8.46 (1H, s), 9.37 (1H, s), 9.43(1H, s), 10.57 (1H, s) V-132 497 / 1.52-1.66 (2H, m), 1.67-1.88 (4H, m),2.00-2.14 (2H, m), 3.29 (3H, s), 3.65 (2H, d), 4.89 (1H, t), 7.58 (1H,d), 7.65 (1H, t), 7.82-7.86 (1H, m), 7.90 (1H, d), 7.98 (1H, d), 8.10(1H, d), 8.79 (1H, s), 9.33 (1H, s), 9.41 (1H, s), 10.55 (1H, s) V-133443 / 3.37-3.45 (2H, m), 3.52-3.65 (2H, m), 4.82 (1H, t), 7.58 (1H, d),7.68 (1H, t), 7.91 (1H, d), 7.98 (1H, d), 8.34 (1H, dd), 8.60 (1H, t),8.78 (1H, s), 8.94 (1H, t), 9.40 (1H, s), 10.50 (1H, s) V-134 457 / 3.29(3H, s), 3.30 (3H, s), 3.45-3.58 (4H, m), 7.58 (1H, d), 7.68 (1H, t),7.91 (1H, d), 7.98 (1H, d), 8.33 (1H, dd), 8.65 (1H, t), 8.77 (1H, s),8.93 (1H, s), 9.40 (1H, s), 10.50 (1H, s) V-135 484 / 1.83 (3H, s),3.22-3.32 (5H, m), 3.32-3.42 (2H, m), 7.56-7.62 (1H, m), 7.70 (1H, t),7.91 (1H, d), 7.98 (1H, d), 8.05 (1H, t), 8.41 (1H, dd), 8.72 (1H, t),8.75 (1H, s), 8.84 (1H, t), 9.41 (1H, s), 10.50 (1H, s) V-136 482 /1.95-2.24 (2H, m), 3.27 (3H, s), 3.54-4.68 (8H, m), 7.57 (1H, d), 7.64(1H, t), 7.91-7.99 (2H, m), 8.02 (1H, d), 8.44 (1H, s), 8.80 (1H, s),9.28 (1H, s), 9.43 (1H, s), 10.56 (1H, s) V-137 496 / 0.80-1.23 (6H, m),2.72-2.93 (2H, m), 3.25 (3H, s), 3.82-4.61 (2H, m), 7.56 (1H, d), 7.62(1H, t), 7.92 (1H, d), 7.96 (1H, dd), 8.01 (1H, d), 8.34 (1H, s), 9.35(1H, s), 9.42 (1H, s), 10.55 (1H, s) V-138 524 / 1.70-1.81 (1H, m),1.81-1.95 (2H, m), 1.94-2.12 (2H, m), 3.25 (3H, s), 3.48-3.62 (2H, m),3.62-3.92 (5H, m), 3.92-4.07 (1H, m), 7.51-7.60 (1H, m), 7.63 (1H, t),7.86-7.96 (1H, m), 7.96-8.07 (2H, m), 8.39 (1H, s), 9.04-9.34 (1H, m),9.41 (1H, d), 10.53 (1H, s) V-139 471 / 3.25 (6H, s), 3.38-3.53 (2H, m),3.59 (2H, t), 3.64-4.06 (3H, m), 7.56 (1H, dt), 7.62 (1H, t), 7.77-7.94(2H, m), 7.98 (1H, d), 8.43 (1H, s), 9.40 (2H, s), 10.53 (1H, s) V-140455 / 1.22 (3H, s), 1.23 (3H, s), 3.06-3.29 (3H, m), 3.23 (3H, s), 4.71(1H, m), 7.56 (1H, d), 7.62 (1H, t), 7.74-7.95 (1H, m), 7.91 (1H, d),7.99 (1H, d), 8.28-8.55 (1H, m), 9.41 (1H, s), 9.42-9.71 (1H, m), 10.54(1H, s) V-141 427 / 2.97-3.21 (3H, m), 3.26 (3H, s), 3.37-3.54 (3H, m),7.56 (1H, dt), 7.62 (1H, t), 7.86 (1H, ddd), 7.92 (1H, d), 7.99 (1H, d),8.45 (1H, s), 9.41 (2H, s), 10.55 (1H, s) V-142 483 / 1.84-2.19 (4H, m),3.25 (3H, s), 3.41-3.55 (1H, m), 3.58-3.71 (1H, m), 3.89-4.01 (1H, m),4.02-4.14 (1H, m), 4.17-4.32 (1H, m), 7.54-7.59 (1 H, m), 7.62 (1H, t),7.82-7.89 (1H, m), 7.92 (1H, d), 7.99 (1H, d), 8.52 (1H, s), 9.30 (1H,s), 9.41 (1H, s), 10.54 (1H, s) V-143 497 / 1.03-1.29 (6H, m), 3.25 (3H,s), 3.47-3.75 (2H, m), 3.97-4.63 (2H, m), 7.54-7.59 (1H, m), 7.62 (1H,t), 7.88-7.96 (2H, m), 8.01 (1H, d), 8.37 (1H, s), 9.39 (1H, s), 9.42(1H, s), 10.56 (1H, s) V-144 510 / 0.78-0.87 (1H, m), 1.14-1.31 (2H, m),1.50-1.69 (2H, m), 1.74-1.90 (2H, m), 3.26 (3H, s), 6.86 (1H, s), 7.35(1H, s), 7.55 (1H, d), 7.63 (1H, t), 7.91 (1H, d), 7.94-8.04 (2H, m),8.35 (1H, s), 9.30 (1H, s), 9.41 (1H, s), 10.54 (1H, s), V-145 511 /3.23 (2H, t), 3.26-3.32 (2H, m), 3.30 (3H, s), 3.40-3.51 (4H, m), 6.32(1H, s), 7.58 (1H, ddd), 7.69 (1H, t), 7.91 (1H, d), 7.98 (1H, d), 8.38(1H, dd), 8.67-8.76 (2H, m), 8.84 (1H, s), 9.40 (1H, s), 10.49 (1H, s)V-146 482 / 1.66-1.80 (1H, m), 2.01-2.19 (1H, m), 2.54-2.65 (1H, m),2.89-3.03 (1H, m), 3.09-3.23 (1H, m), 3.22-3.55 (4H, m), 3.30 (3H, s),7.59 (1H, d), 7.67 (1H, t), 7.92 (1H, d), 7.99 (1H, d), 8.21 (1H, dd),8.71-8.91 (3H, m), 9.04 (1H, s), 9.42 (1H, s), 10.53 (1H, s) V-147 510 /2.05 (3H, s), 3.25 (3H, s), 3.51-3.67 (4H, m), 3.68-3.98 (4H, m), 7.57(1H, d), 7.63 (1H, t), 7.89 (1H, d), 7.93 (1H, d), 8.01 (1H, d), 8.41(1H, s), 9.39 (1H, s), 9.42 (1H, s), 10.56 (1H, s) V-148 510 / 1.84 (3H,s), 1.86-2.06 (2H, m), 2.07-2.28 (2H, m), 3.25 (3H, s), 3.26 (3H, s),3.41-3.52 (1H, m), 3.57-3.86 (4H, m), 4.05-4.19 (2H, m), 4.23-4.33 (2H,m), 4.33-4.43 (1H, m), 7.54-7.59 (2H, m), 7.59-7.67 (2H, m), 7.81-7.96(2H, m), 7.91 (2H, d), 7.98 (2H, d), 8.19 (2H, t), 8.45 (1H, s), 8.54(1H, s), 9.25 (1H, s), 9.30 (1H, s), 9.40 (2H, s), 10.54 (2H, s) V-149496 / 1.26-1.49 (2H, m), 1.84-2.00 (3H, m), 2.76-2.96 (2H, m), 3.14-3.39(5H, m), 7.56 (1H, d), 7.66 (1H, t), 7.89 (1H, d), 7.94 (1H, d), 8.08(1H, dd), 8.62 (1H, t), 8.73 (1H, s), 9.14 (1H, s), 9.35 (1H, s) V-150483 / 1.54-1.70 (2H, m), 1.86 (2H, dd), 3.31 (3H, s), 3.39-3.46 (2H, m),3.86-3.96 (2H, m), 3.96-4.11 (1H, m), 7.59 (1H, d), 7.66 (1H, t), 7.90(1H, d), 7.98 (1H, d), 8.11 (1H, dd), 8.38 (1H, d), 8.77 (1H, s), 9.18(1H, s), 9.40 (1H, s), 10.52 (1H, s) V-151 497 / 1.19-1.34 (2H, m),1.34-1.52 (2H, m), 1.82-1.98 (4H, m), 3.30 (3H, s), 3.39-3.51 (1H, m),3.63-3.85 (1H, m), 4.56-4.64 (1H, m), 7.59 (1H, d), 7.66 (1H, t), 7.91(1H, d), 7.98 (1H, d), 8.12 (1H, d), 8.24 (1H, d), 8.75 (1H, s), 9.18(1H, s), 9.41 (1H, s), 10.53 (1H, s) V-152 457 / 2.99-3.12 (1H, m), 3.22(3H, s), 3.39-3.51 (1H, m), 3.56-3.60 (1H, m), 3.61-3.69 (3H, m),3.69-3.86 (1H, m), 7.54 (1H, d), 7.58-7.70 (1H, m), 7.81-7.88 (1H, m),7.89 (1H, d), 7.96 (1H, d), 8.36-8.58 (1H, m), 9.29-9.37 (1H, m), 9.38(1H, s) V-153 471 / 1.22 (3H, d), 3.30 (3H, s), 3.31 (3H, s), 3.33-3.40(1H, m), 3.47 (1H, dd), 4.14-4.30 (1H, m), 7.56-7.61 (1H, m), 7.66 (1H,t), 7.91 (1H, d), 7.98 (1H, d), 8.12 (1H, ddd), 8.26 (1H, d), 8.78 (1H,s), 9.21 (1H, s), 9.40 (1H, s), 10.53 (1H, s) V-154 483 / 1.36-1.55 (2H,m), 1.75-1.95 (2H, m), 3.25 (3H, s), 3.41-3.60 (2H, m), 3.72-3.89 (1H,m), 3.95-4.13 (2H, m), 4.73-4.98 (1H, m), 7.56 (1H, td), 7.63 (1H, t),7.91 (1H, d), 7.93-7.98 (1H, m), 8.00 (1H, d), 8.35 (1H, s), 9.32 (1H,s), 9.41 (1H, s), 10.54 (1H, s), V-155 497 / 1.84-2.02 (2H, m),2.02-2.16 (1H, m), 3.25 (3H, s), 3.29 (3H, s), 3.37-3.49 (2H, m),3.50-3.67 (1H, m), 3.86-4.01 (1H, m), 4.01-4.18 (1H, m), 4.28-4.46 (1H,m), 7.54-7.59 (1H, m), 7.62 (1H, t), 7.78-7.88 (1H, m), 7.91 (1H, d),7.98 (1H, d), 8.51 (1H, s), 9.28-9.36 (1H, m), 9.40 (1H, s), 10.53 (1H,s), V-156 469 / 1.87-2.02 (1H, m), 2.02-2.18 (1H, m), 3.60-3.74 (1H, m),3.21 (3H, s), 3.76-4.14 (2H, m), 4.34-4.51 (1H, m), 4.52-4.97 (1H, m),7.49-7.66 (2H, m), 7.80-7.95 (2H, m), 8.00 (1H, d), 8.51 (1H, s), 9.13(1H, s), 9.35 (1H, s), 9.89-10.28 (1H, m) V-157 498 / 1.27 (3H, t), 2.88(6H, s), 3.25 (3H, s), 3.31-3.48 (2H, m), 3.61-3.80 (2H, m), 3.74-4.18(2H, m), 7.57 (1H, td), 7.62 (1H, t), 7.93 (1H, d), 8.01 (1H, d),8.04-8.15 (1H, m), 8.42 (1H, s), 9.04-9.26 (0.5H, m), 9.43 (1H, s),9.48-9.69 (0.5H, m), 10.54 (1H, s) V-158 410 / (CDCl₃) 0.98-1.11 (2H,m), 1.13-1.34 (3H, m), 1.60-1.79 (2H, m), 1.80-1.89 (2H, m), 1.89-2.03(2H, m), 2.97 (4H, t), 3.44 (2H, t), 3.83 (4H, t), 5.47 (1H, t), 7.55(2H, s), 8.21 (1H, s), 8.95 (1H, s) V-159 452 / (CDCl₃) 2.88-3.05 (6H,m), 3.75-3.89 (6H, m), 5.48 (1H, t), 7.19 (2H, d), 7.27 (2H, d), 7.56(1H, d), 7.59 (1H, d), 8.21 (1H, s), 8.95 (1H, s) V-160 510 / 1.90-2.03(2H, m), 2.03-2.20 (2H, m), 2.78 (2H, s), 3.03-3.31 (6H, m), 3.26 (3H,s), 3.43-3.62 (2H, m), 4.41-4.69 (1H, m), 7.54-7.59 (1H, m), 7.62 (1H,t), 7.77-7.90 (1H, m), 7.93 (1H, d), 8.01 (1H, d), 8.47 (1H, s), 9.43(1H, s), 9.44-9.65 (1H, m), 10.58 (1H, s) V-161 382 / (CDCl₃) 1.50-1.62(2H, m), 1.62-1.83 (4H, m), 2.08-2.21 (2H, m), 2.90-3.00 (4H, m),3.77-3.84 (4H, m), 4.40-4.52 (1H, m), 5.40 (1H, d, 7.3 Hz), 7.55 (2H,s), 8.21 (1H, s), 8.95 (1H, s) V-162 452 / (CDCl₃) 2.91-3.03 (4H, m),3.17 (2H, t), 3.81-3.87 (4H, m), 3.90 (2H, td), 5.56 (1H, t), 7.12-7.22(2H, m), 7.28 (1H, dd), 7.37 (1H, dd), 7.57 (1H, d), 7.60 (1H, d), 8.26(1H, s), 8.96 (1H, s) V-163 432 / (CDCl₃) 1.40 (3H, d), 2.89-3.03 (4H,m), 3.08-3.30 (1H, m), 3.63-3.74 (1H, m), 3.79-3.86 (4H, m), 3.86-3.96(1H, m), 5.37 (1H, t), 7.20-7.40 (5H, m), 7.56 (1H, d), 7.59 (1H, d),8.24 (1H, s), 8.94 (1H, s) V-164 468 / (CDCl₃) 1.78 (3H, d), 2.83 (4H,s), 3.67 (4H, s), 5.91 (1H, s), 6.12-6.24 (1H, m), 7.42 (1H, dd),7.48-7.60 (4H, m), 7.65 (1H, d), 7.74 (1H, d), 7.78-7.92 (1H, m), 7.89(1H, d), 8.32 (1H, d), 8.95 (1H, s) V-165 433 / (MeOD) 2.86 (2H, t),2.94-3.06 (4H, m), 3.67-3.74 (2H, m), 3.81-3.91 (4H, m), 6.69 (2H, d),7.05 (2H, d), 7.65 (1H, d), 7.68 (1H, d), 8.25 (1H, s), 8.99 (1H, s)V-166 473 / 2.26 (3H, s), 2.74 (4H, s), 3.10 (2H, t), 3.70 (4H, s), 4.14(2H, t), 7.14 (2H, s), 7.84 (1H, d), 7.92 (1H, d), 8.75 (1H, s), 9.30(1H, s), 9.67 (1H, s), 10.03 (1H, s) V-167 452 / 2.96 (2H, s), 3.23 (4H,s), 3.70 (2H, t), 3.93 (4H, s), 7.45-7.08 (4H, m), 7.77 (2H, s), 8.24(1H, s), 9.10 (2H, s) V-168 438 / 3.24 (4H, s), 3.93 (4H, s), 4.67 (2H,d), 7.23-7.29 (1H, m), 7.33 (1H, t), 7.44 (1H, s), 7.53 (1H, s), 7.78(2H, d), 8.16-8.36 (1H, m), 9.03 (1H, s), 9.16 (1H, s) V-169 424 /0.92-1.27 (7H, m), 1.43-1.87 (6H, m), 3.24 (4H, s), 3.90 (4H, s),4.03-4.26 (1H, m), 7.49 (1H, d), 7.74 (2H, s), 8.19 (1H, s), 8.91 (1H,s), 9.03-9.24 (1H, m) V-170 432 / 1.93 (2H, s), 2.69 (2H, t), 3.23 (4H,s), 3.49 (2H, t), 3.92 (4H, s), 7.05-7.39 (5H, m), 7.77 (2H, s), 8.16(1H, s), 9.09 (2H, s) V-171 407 / (CDCl₃) 0.82-0.96 (1H, m), 1.17-1.34(1H, m), 1.34-1.47 (1H, m), 1.50-1.76 (2H, m), 2.14-2.34 (2H, m),2.62-2.86 (3H, m), 2.90-3.03 (4H, m), 3.74-3.90 (4H, m), 4.23-4.42 (1H,m), 5.68 (2H, d), 7.54 (2H, s), 8.22 (1H, s), 8.93 (1H, s) V-172 430 /2.68-2.85 (5H, m), 3.00 (2H, dd), 3.64 (4H, s), 4.76-4.96 (1H, m), 7.15(2H, dd), 7.18-7.30 (2H, m), 7.76 (2H, s), 7.96 (1H, s), 8.11 (1H, s),9.15 (1H, s) V-173 419 / 2.85 (4H, s), 2.97 (2H, s), 3.62-3.78 (6H, m),7.19-7.38 (1H, m), 7.64-7.82 (4H, m), 8.16 (1H, s), 8.37 (1H, dd), 8.51(1H, s), 9.11 (1H, s) V-174 482 / 2.75-2.90 (4H, m), 3.14 (3H, s),3.62-3.79 (4H, m), 4.74 (2H, d), 7.59-7.80 (4H, m), 7.85 (2H, d), 8.09(1H, s), 8.36 (1H, s), 9.16 (1H, s) V-175 448 / 2.88 (6H, s), 3.54-3.67(3H, m), 3.68-3.78 (6H, m), 6.85 (2H, d), 7.21 (2H, d), 7.69 (1H, s),7.75 (2H, s), 8.17 (1H, s), 9.12 (1H, s) V-5 418 / 2.32 (6H, s), 3.26(4H, s), 3.99 (4H, s), 6.68 (1H, s), 7.69 (2H, s), 7.89 (1H, d), 7.95(1H, d), 8.27 (1H, s), 8.93 (1H, s), 9.34 (1H, s), 9.95 (1H, s) V-178408 / (CD₃OD) 2.65 (1H, s), 3.16 (2H, t), 3.42-3.33 (4H, m), 3.98 (2H,t), 4.18-4.05 (4H, m), 7.38 (1H, s), 7.83 (2H, s), 8.47 (1H, s), 8.76(1H, s), 9.21 (1H, s) V-179 419 / (CD₃OD) 3.03 (4H, d), 3.22-3.10 (2H,m), 3.97-3.80 (6H, m), 7.25-7.13 (1H, m), 7.36 (1H, d), 7.79-7.51 (3H,m), 8.27-8.17 (1H, m), 8.44 (1H, d), 8.95 (1H, d) V-180 436 / (CD₃OD)3.00 (2H, t), 3.44-3.33 (4H, m), 3.84 (2H, s), 4.18-4.00 (4H, m), 6.99(2H, t), 7.29 (2H, dd), 7.75 (2H, s), 8.38 (1H, s), 9.18 (1H, s) V-181394 / (CD₃OD) 3.43-3.33 (4H, m), 4.14-3.93 (6H, m), 5.96 (1H, s), 6.39(2H, d), 7.22 (1H, d), 7.70 (2H, d), 8.07 (1H s) V-182 434 / (CD₃OD)2.90 (2H, t), 3.43-3.32 (4H, m), 3.77 (2H, s), 4.15-4.02 (4H, m), 6.70(2H, d), 7.10 (2H, d), 7.70 (2H, s), 8.32 (1H, s), 9.08 (1H, s)

Example 26

Thiophene-3-carboxylic acid methoxymethyl amide

Thiophene-3-carboxylate (10 g, 78 mmol) was dissolved in chloroform (500ml), treated with thionyl chloride (6.83 ml, 93.6 mmol) and heated toreflux for 2 hours. This solution was then added by cannulation to asolution of Weinreb Amine (11.4 g, 117 mmol) and triethylamine (30 ml,210.6 mmol) in chloroform (100 ml). The reaction was then stirred atroom temperature overnight. The solvent was reduced in vacuo to 100 ml,washed with water (150 ml), dried (Na₂SO₄) and concentrated. Theresulting orange oil was purified by vacuum distillation (150° C. at 5mbar) to give the title compound as a pale yellow oil (8.50 g, 64%). MS(ES⁺) 172. δH (d⁶ DMSO): 3.25 (3H, s), 3.64 (3H, s), 7.43 (1H, d), 7.56(1H, d), 8.25 (1H, s).

Example 27

4-Methyl-1-thiophen-3-yl-pentan-1-one

Magnesium powder (1.49 g, 61.3 mmol) was added to dry diethyl ether (100ml) under nitrogen atmosphere. One iodine crystal (catalytic) was added,followed by 1-bromo-3-methylbutane (6.3 ml, 52.6 mmol). The reaction wasinitiated by sonication at 30° C. and was allowed to spontaneouslyreflux. Once the exothermic reaction had subsided, the mixture was addedby cannulation to a solution of thiophene-3-carboxylic acidmethoxy-methyl-amide (7.50 g, 43.8 mmol) in dry diethyl ether (10 ml).Once addition was complete, the mixture was stirred at room temperaturefor 2 hours. The reaction mixture was then washed with saturatedammonium chloride (2×150 ml), dried (Na₂SO₄) and concentrated in vacuo.The crude yellow oil was purified by silica gel chromatography to yieldthe title compound as a colourless oil (1.55 g, 19%). 8H (d⁶ DMSO): 0.88(6H, d), 1.49 (2H, q), 1.56 (1H, m), 2.90 (2H, t), 7.49 (1H, d), 7.59(1H, d), 8.50 (1H, s).

Example 28

(4-Methyl-1-thiophen-3-yl-pentylideneaminooxy)-acetic acid

4-Methyl-1-thiophen-3-yl-pentan-1-one (324 mg, 1.78 mmol) was dissolvedin dry methanol (20 ml) under nitrogen atmosphere, treated withcarboxymethoxylamine hemihydrochloride (194 mg, 1.78 mmol) and sodiumhydroxide (35 mg, 0.89 mmol) and stirred at room temperature for 4hours. The solvent is then removed in vacuo and the resulting oil wastriturated with hexane to give the title compound as a white solid (307mg, 67%). MS (ES⁺) 25. δH (d⁶ DMSO): 1.00 (6H, d), 1.45 (2H, q), 1.63(1H, m), 2.75 (2H, t), 4.65 (2H, s), 7.39 (1H, d), 7.60 (1H, d), 7.86(1H, s), 12.70 (1H, s).

Example 29

7,7-Dimethyl-6,7-dihydro-5H-benzo[b]thiophen-4-one

(4-Methyl-1-thiophen-3-yl-pentylideneaminooxy)-acetic acid (2.17 g, 8.5mmol) was added to a 0.1 M sodium hydroxide solution (93.5 ml) andheated to reflux. A solution of potassium persulfate (3.45 g, 12.75mmol) in water (10 ml) was added dropwise and heating continued. After 2hours, the reaction mixture was extracted with dry diethyl ether (100ml). The organic layer was then washed with saturate sodium bicarbonate,water and brine, dried (Na₂SO₄) and concentrated in vacuo. The resultingoil was purified by silica gel chromatography to afford the titlecompound as a colourless oil (196 mg, 13%). MS (ES⁺) 18. δH (d⁶DMSO):1.42 (6H, s), 2.03 (2H, t), 2.60 (2H, t), 7.22 (1H, d), 7.42 (1H, d).

Example 30

3-(6,6-Dimethyl-5,6-dihydrothieon[2,3-h]quinazolin-2-ylamino)-benzenesulfonamide

7,7-Dimethyl-6,7-dihydro-5H-benzo[b]thiophen-4-one (196 mg, 1.09 mmol)was dissolved in DME (5 ml), treated with Brederick's reagent (336 μl,1.63 mmol) and heated to reflux for 12 hours. The solvent was thenremoved in vacuo to give an orange solid which was triturated withpentane to afford the enaminone as a yellow solid (141 mg, 55%). MS(ES⁺) 236.2. δH (d⁶ DMSO): 0.80-0.88 (6H, m), 2.87 (2H, s), 3.08 (6H,s), 7.19 (1H, d), 7.28 (1H, d), 7.48 (1H, s).

3-Guanidino-benzenesulfonamide hydrochloride (225 mg, 0.9 mmol) wasdissolved in DMA (10 ml), treated with potassium carbonate (62 mg, 0.45mmol) and heated to 50° C. for 10 minutes.5-Dimethylaminomethylene-7,7-dimethyl-6,7-dihydro-5H-benzo[b]thiophen-4-one(141 mg, 0.6 mmol) was the added and the reaction mixture heated toreflux for 12 hours. The mixture was extracted in ethyl acetate (100 ml)and washed with water (2×100 ml), dried (Na₂SO₄) and concentrated togive an orange oil. This was initially purified by flash chromatographyand then by reverse phase preparative HPLC [Waters Delta-Pak C18, 15 uM,100 A column, gradient 10%-100% B (solvent A: water; solvent B: CH₃CN)over 10 minutes at 25 mL/min] to afford the title compound as a whitesolid (22 mg, 9%). MS (ES⁺) 385.2. δH (d⁶ DMSO): 1.30 (6H, s), 2.85 (2H,s), 7.30 (2H, s), 7.36 (1H, d), 7.45 (1H, t), 7.53 (1H, d), 7.66 (1H,d), 7.77 (1H, d), 8.39 (1H, s), 8.78 (1H, s), 9.90 (1H, s).

Example 31

3-(8-Hydroxy-5,6-dihydro-quinazolin-2-ylamino)benzenesulfonamide

A mixture of 2-(tert-butyl-dimethyl-silanyloxy)-cyclohex-2-enone (6.9 g,30.48 mmol) in dry toluene (50 mL) was treated with Bredereck's reagent(9.44 ml, 45.72 mmol) was stirred at 115° C. for 24 hours. The mixturewas concentrated under reduced pressure to give2-(tert-butyl-dimethyl-silanyloxy)-6-dimethylaminomethylene-cyclohex-2-enoneas a semi-solid brown residue which was used as such in the next step.

The crude2-(tert-butyl-dimethyl-silanyloxy)-6-dimethylaminomethylene-cyclohex-2-enonein DMA (75 mL) was treated with 3-guanidino-benzenesulfonamide as thehydrochloride salt (7.79 g, 31.09 mmol) and potassium carbonate (2.13 g,15.54 mmol) and the mixture stirred at 110° C. for 20 hours. The mixturewas evaporated under high vacuum and triturated with dichloromethane(100 mL) to give a buff coloured precipitate. The precipitate wasfiltered, the mother liquors concentrated in vacuo and purified by flashchromatography on silica gel eluting with ethyl acetate to give3-(8-hydroxy-5,6-dihydro-quinazolin-2-ylamino)benzenesulfonamide as ayellow solid (0.63 g, 6.5%). MS (ES⁺) 319, (ES⁻) 317. δH (d⁶ DMSO)2.04-2.12 (2H, m), 2.66-2.74 (2H, m), 2.84-2.90 (2H, m), 7.33 (2H, s),7.43 (1H, d), 7.48 (1H, t), 8.16 (1H, d), 8.25 (1H, s), 8.78 (1H, s),10.25 (1H, s).

Example 32

3-(2-Methyl-4,5-dihydro-3-thia-1,7,9-triaza-cyclopenta[α]naphthalen-8-ylamino)-benzenesulfonamide

A solution of3-(8-hydroxy-5,6-dihydro-quinazolin-2-ylamino)benzenesulfonamide (100mg, 0.314 mmol) in chloroform (1 mL) was treated with a solution ofbromine (32 μl, 0.628 mmol) in chloroform (1 mL) and the mixture stirredat room temperature for 2 hours giving an insoluble black residue. Thesolvent was decanted and the residue treated with thioacetamide (47 mg,0.6 mmol) in ethanol (3 mL). The mixture was stirred at 80° C. for 3hours and filtered to give a black solid (75 mg). The solid wasdissolved in DMSO and purified by reverse phase preparative HPLC [WatersDelta-Pak C18, 15 uM, 100 A column, gradient 10%-100% B (solvent A:0.05% TFA in water; solvent B: CH₃CN) over 10 minutes at 25 mL/min] togive a yellow solid (6.9 mg) which was further purified by flashchromatography on silica gel eluting with ethyl acetate to afford thetitle compound (1.9 mg) as a pale yellow solid. MS (ES⁺) 374, (ES⁻) 372.δH (CD₃OD) 2.78 (3H, s), 3.01-3.07 (2H, m), 3.10-3.16 (2H, m), 7.46 (1H,t), 7.53 (1H, d), 7.65 (2H, s, partially exchanged), 7.83 (1H, d), 8.27(1H, s), 8.48 (1H, s).

Example 33

3-(2-Methyl-4,5-dihydro-thiazolo[4,5-h]quinazolin-8-ylamino)-benzenesulfonamide

Sodium (76 mg, 3.33 mmol) in diethylether (6 mL) was treated with ethylformate (0.146 mL, 1.80 mmol) and2-methyl-5,6-dihydro-4H-benzothiazol-7-one (0.251 g, 1.50 mmol). Thereaction mixture was cooled down to 0° C. and ethanol (0.193 mL, 3.30mmol) was added. The cooling bath was removed and the reaction mixturewas stirred at room temperature for 4 hours. The solvent was thenremoved in vacuo and the resulting crude product was taken through tothe next step without further purification.

The above was dissolved in anhydrous DMA (5 mL) and treated with3-guanidino-benzenesulfonamide hydrogen chloride (384 mg, 1.53 mmol) andpowdered potassium carbonate (106 mg, 0.77 mmol). The mixture was heatedto 120° C. with vigorous stirring for 12 hours. The mixture was cooleddown to room temperature, extracted in ethyl acetate (20 ml) and washedwith water (2×20 ml), dried (MgSO₄) and concentrated in vacuo. This wasinitially purified by flash chromatography and then by reverse phasepreparative HPLC [Waters Delta-Pak C18, 15 uM, 100 A column, gradient10%-100% B (solvent A: 0.05% TFA in water; solvent B: CH₃CN) over 10minutes at 25 mL/min] to afford the title compound (20 mg) as a yellowpowder. MS (ES⁺) 374, (ES⁻) 372. δH (d⁶ DMSO) 2.73 (3H, s), 3.00 (4H,m), 7.31 (2H, s), 7.38 (1H, s), 7.46 (1H, t), 7.89 (1H, d), 8.36 (1H,s), 8.41 (1H, s), 9.91 (1H, s).

A variety of other compounds of formula III have been prepared bymethods substantially similar to those described in Example 33. Thecharacterization data for these compounds is summarized in Table 16below and includes HPLC, LC/MS (observed) and ¹H NMR data.

¹H NMR data is summarized in Table 16 below wherein ¹H NMR data wasobtained at 400 MHz in deuterated DMSO, unless otherwise indicated, andwas found to be consistent with structure. Compound numbers correspondto the compound numbers listed in Table 3.

TABLE 16 Characterization Data for Selected Compounds of Formula IIICom- pound M + 1 No. (obs) Rt (min) ¹H-NMR III-31 474 8.9 1.22 (9H, s),2.99 (2H, d), 3.05 (2H, d), 5.42 (2H, s), 7.28 (2H, s), 7.37 (1H, d),7.44 (1H, t), 7.83 (1H, d), 8.39 (1H, s), 8.43 (1H, s), 9.93 (1H, s)III-32 445 7.5 2.87 (4H, m), 3.54 (4H, t), 3.74 (4H, t), 7.29 (2H, s),7.35 (1H, m), 7.43 (1H, t), 7.89 (1H, m), 8.16 (1H, s), 8.41 (1H, t),9.73 (1H, s) III-33 431 8.4 1.21 (6H, t), 2.85 (4H, m), 3.54 (4H, q),7.28 (2H, s), 7.35 (1H, m), 7.43 (1H, t), 7.87 (1H, m), 8.12 (1H, s),8.42 (1H, t), 9.66 (1H, s) III-34 443 8.7 1.63 (6H, s), 2.84 (4H, m),3.56 (4H, s), 7.29 (2H, s), 7.35 (1H, d), 7.43 (1H, t), 7.89 (1H, d),8.13 (1H, s), 8.42 (1H, s), 9.69 (1H, s) III-35 472 7.1 1.44-1.54 (2H,m), 1.89-2.10 (4H, m), 2.16 (3H, s), 2.67-2.74 (2H, m), 2.76-2.88 (4H,m), 3.58 (1H, br s), 7.27 (2H, s), 7.35 (1H, m), 7.42 (1H, t), 7.89 (1H,m), 8.10 (1H, s), 8.37-8.40 (2H, m), 9.64 (1H, s) III-36 444 7.02.75-2.90 (8H, m), 3.47 (4H, t), 7.29 (2H, s), 7.35 (1H, d), 7.43 (1H,t), 7.89 (1H, d), 8.14 (1H, s), 8.41 (1H, s), 9.71 (1H, s) III-37 4867.1 2.06 (3H, s), 2.80-2.93 (4H, m), 3.50-3.78 (8H, m), 7.29 (2H, s),7.36 (1H, d), 7.44 (1H, t), 7.89 (1H, d), 8.16 (1H, s), 8.40 (1H, t),9.72 (1H, s) III-38 390 6.4 2.95-3.05 (4H, m), 4.76 (2H, d), 6.22 (1H,t), 7.29 (2H, s), 7.39 (1H, d), 7.48 (1H, t), 7.94 (1H, d), 8.39 (2H,s), 9.90 (1H, s) III-39 458 7.7 2.24 (3H, s), 2.44 (4H, t), 2.86 (4H,m), 3.55 (4H, t), 7.29 (2H, s), 7.35 (1H, m), 7.43 (1H, t), 7.89 (1H,m), 8.15 (1H, s), 8.40 (1H, t), 9.72 (1H, s) III-40 513 8.1 2.24 (3H,s), 2.44 (4H, t), 2.86 (4H, m), 2.92 (3H, s), 3.15 (3H, s), 3.55 (4H,t), 7.26 (1H, m), 7.39 (1H, t), 7.92 (1H, dd), 8.15 (1H, s), 8.20 (1H,s), 8.30 (1H, t), 9.68 (1H, s)

Example 34

7-Oxo-4,5,6,7-tetrahydro-benzothiazole-2-carboxylic acid ethyl ester

A mixture of 2-bromo-cyclohexane-1,3-dione (3.84 g, 20.10 mmol) andethyl thiooxamate (1.339 g, 10.05 mmol) in pyridine (20 mL) was heatedovernight at 50° C. The crude reaction mixture was concentrated invacuo, taken up in dichloromethane and washed with water and brine. Theorganic extract was dried over magnesium sulfate, filtered andconcentrated in vacuo. The residue was purified by silica gelchromatography eluting with EtOAc:hexanes (30:70) to give the titlecompound in 20% yield (0.45 g). MS (ES⁺) 226. δH (CDCl₃) 1.47 (3H, t),2.28 (2H, quint.), 2.71 (2H, t), 3.18 (2H, t), 4.53 (2H, q).

Example 35

8-(3-Sulfamoyl-phenylamino)-4,5-dihydro-thiazolo[4,5-h]quinazoline-2-carboxylicacid ethyl ester

To 7-oxo-4,5,6,7-tetrahydro-benzothiazole-2-carboxylic acid ethyl ester(0.1128 g, 0.50 mmol) was added DMF/DMA solution (3 mL) and the mixturewas refluxed under nitrogen for 1 hour. The reaction mixture wasconcentrated in vacuo and the resulting crude product(6-dimethylaminomethylene-7-oxo-4,5,6,7-tetrahydro-benzothiazole-2-carboxylicacid ethyl ester) was taken through to the next step without furtherpurification. The above was dissolved in anhydrous DMA (3 mL) andtreated with 3-guanidino-benzenesulfonamide hydrogen chloride (128 mg,0.51 mmol) and powdered potassium carbonate (35 mg, 0.26 mmol). Themixture was heated to 120° C. with vigorous stirring for 12 hours. Themixture was cooled down to room temperature, and diluted with water. Theresulting solid was filtered, rinsed with more water, a little bit of^(i)PrOH and Et₂O. The crude solid was purified by silica gel to givethe title compound as a brown solid in 24% yield (52 mg). MS (ES⁺) 432,(ES⁻) 430. δH (d⁶ DMSO) 1.36 (3H, t), 3.05 (2H, t), 3.17 (2H, t), 4.42(2H, q), 7.33 (2H, s), 7.41 (1H, d), 7.50 (1H, t), 7.89 (1H, d), 8.43(1H, t), 8.52 (1H, s), 10.04 (1H, s).

Example 36

8-(3-Sulfamoyl-phenylamino)-thiazolo[4,5-h]quinazoline-2-carboxylic acidethyl ester

A suspension of8-(3-Sulfamoyl-phenylamino)-4,5-dihydro-thiazolo[4,5-h]quinazoline-2-carboxylicacid ethyl ester (609 mg, 1.41 mmol) and2,3-dichloro-5,6-dicyano-1,4-benzoquinone (480 mg, 2.12 mmol) inanhydrous 1,4-dioxane (30 mL) was refluxed under nitrogen for 2 hours.The reaction mixture was cooled down to room temperature and the DDQresidues were filtered. The mother liquor was concentrated in vacuo andthe resulting crude residue was triturated in a mixture of EtOAc, MeOHand DCM. The resulting solid was filtered to afford 523 mg (86% yield)of the title compound as a yellow solid. MS (ES⁺) 430, (ES⁻) 428. δH (d⁶DMSO) 1.42 (3H, t), 4.51 (2H, q), 7.37 (2H, s), 7.52 (1H, d), 7.60 (1H,t), 8.09 (1H, d), 8.14 (2H, s), 8.70 (1H, s), 9.55 (1H, s), 10.59 (1H,s).

Example 37

8-(3-Sulfamoyl-phenylamino)-thiazolo[4,5-h]quinazoline-2-carboxylic acid

8-(3-Sulfamoyl-phenylamino)-thiazolo[4,5-h]quinazoline-2-carboxylic acidethyl ester (494 mg, 1.15 mmol) was suspended in a mixture of EtOH (30ml) and 1N sodium hydroxide (20 ml). The mixture was stirred at roomtemperature for 12 h. EtOH was removed in vacuo and the residual crudemixture was neutralised with 1N HCl (20 ml). The resulting solid wasfiltered and dried under vacuo to afford 0.473 g of a brownish solid. MS(ES⁺) 402, (ES⁻) 400. δH (d⁶ DMSO) 7.37 (2H, s), 7.50 (1H, d), 7.60 (1H,t), 8.06 (2H, s), 8.16 (1H, d), 8.62 (1H, s), 9.50 (1H, s), 10.52 (1H,s), 14.80 (1H, br s).

Example 38

{1-[8-(3-Sulfamoyl-phenylamino)-thiazolo[4,5-h]quinazoline-2-carbonyl]-pyrrolidin-3-yl}-carbamicacid tert-butyl ester

Diisopropylethylamine (68 μl, 0.39 mmol) was added to a mixture of8-(3-Sulfamoyl-phenylamino)-thiazolo[4,5-h]quinazoline-2-carboxylic acid(78 mg, 0.19 mmol), 3-(tert-butoxycarbonylamino)pyrrolidine (36.2 mg,0.19 mmol) and PyBrop (91 mg, 0.19 mmol) in dimethylformamide (1 ml).The reaction mixture was stirred at room temperature for 12 h. Thereaction was concentrated under reduced pressure, redissolved inisopropanol (˜10 mL) and water was added (˜5 mL). The precipitate formedwas isolated by filtration and washed with water (1×5 mL), dilute NaHSO₄(1×5 mL), water (1×5 mL), saturated Na₂CO₃ (1×5 mL), water (1×5 mL),isopropanol (1×5 mL), diethyl ether (4×5 mL) giving a yellow powder (87mg, 80% yield). MS (ES⁺) 570, (ES⁻) 568.

Example 39

3-[2-(3-aminopyrrolidine-1-carbonyl)-thiazolo[4,5-h]quinazolin-8-ylamino]-benzenesulfonamide

Trifluoroacetic acid (1 ml) was added to a suspension of{1-[8-(3-Sulfamoyl-phenylamino)-thiazolo[4,5-h]quinazoline-2-carbonyl]-pyrrolidin-3-yl}-carbamicacid tert-butyl ester (87 mg, 0.15 mmol) in dichloromethane (1 ml). Thereaction mixture was stirred at room temperature for 1 hour. Thesolution was concentrated under reduced pressure and the residue waspurified by reverse phase preparative HPLC [Waters Delta-Pak C18, 15 uM,100 A column, gradient 10%-100% B (solvent A: 0.05% TFA in water;solvent B: CH₃CN) over 10 minutes at 25 mL/min] to afford the titlecompound as a yellow solid. MS (ES⁺) 470, (ES⁻) 468. δH (d⁶ DMSO)2.0-2.5 (4H, m), 3.70-4.05 (3H, m), 4.31-4.44 (2H, m), 7.37 (2H, s),7.52 (1H, d), 7.60 (1H, t), 8.02-8.13 (6H, m), 8.69 (1H, s), 9.55 (1H,d), 10.57 (1H, s).

A variety of other compounds of formula V have been prepared by methodssubstantially similar to those described in Example 38 and 39. Thecharacterization data for these compounds is summarized in Table 17below and includes HPLC, LC/MS (observed) and ¹H NMR data.

¹H NMR data is summarized in Table 17 below wherein ¹H NMR data wasobtained at 400 MHz in deuterated DMSO, unless otherwise indicated, andwas found to be consistent with structure. Compound numbers correspondto the compound numbers listed in Table 5.

TABLE 17 Characterization Data for Selected Compounds of Formula V Com-pound M + 1 Rt No. (obs) (min) ¹H-NMR V-177 484 6.4 1.50-1.67 (2H, m),2.00-2.15 (2H, 760917 m), 3.00-3.13 (1H, m), 3.30-3.50 (2H, m),4.52-4.58 (1H, m), 5.09-5.15 (1H, m), 7.37 (2H, s), 7.51 (1H, d), 7.59(1H, t), 7.92 (3H, br s), 8.02-8.13 (3H, m), 8.74 (1H, s), 9.55 (1H, s),10.58 (1H, s) V-178 470 / 3.25-3.35 (4H, m), 3.94 (2H, s), A 4.56 (2H,s), 7.34 (2H, s), 7.49 (1H, d), 7.58 (1H, t), 8.02-8.13 (3H, m), 8.73(1H, s), 8.92 (2H, s), 9.56 (1H, s), 10.59 (1H, s)

Example 40

2-Dimethylaminomethylene-cyclohexane-1,3-dione

A MeCN solution of cyclohexane-1,3-dione 2.24 g (0.02 mol) i and DMF-DMA11.9 g (0.1 mol) was stirred at room temperature for 4 h. The solventwas removed under reduced pressure and the product was used for the nextstep with no purification.

1-Methyl-1,3a,5,6,7,7a-hexahydro-indazol-4-one: A MeOH solution ofmethylhydrazine 0.046 g (0.01 mol) and2-dimethylaminomethylene-cyclohexane-1,3-dione 1.67 g (0.01 mol) wasstirred at room temperature for 1 h, and then was stirred at 50° C. for1 h. NMR of crude material indicated the reaction was completed. Workup: The reaction mixture was diluted with ethyl acetate and washed withbrine. The organic phase was dried with MgSO₄, and the solvent wasrotary evaporated. The crude product was filtered through silica gel,and 1.03 g pure product was obtained, the yield is 69%.

5-Hydroxymethylene-1-methyl-1,3a,5,6,7,7α-hexahydro-indazol-4-one: To aMeCN solution of 1-methyl-1,3a,5,6,7,7a-hexahydro-indazol-4-one 1.03 g(0.00686 mol) was added Bredereck's reagent 1.79 g (0.010 mol) and themixture was stirred at 75° C. for 2 h. Thin layer chromatographyindicated starting material to be the major component, hence 3equivalents of DMF-DMA were added and the reaction mixture was stirredat 80° C. overnight. The solvent was evaporated and the crude materialwas used for next step.

3-(7-Methyl-5,7-dihydro-6H-pyrazolo[3,4-h]quinazolin-2-ylamino)-phenol:To the DMF solution of crude5-hydroxymethylene-1-methyl-1,3a,5,6,7,7a-hexahydro-indazol-4-one wasadded 10 eq of N-(3-hydroxy-phenyl)-guanidine, the reaction mixture wasstirred at 100° C. for overnight, LC/MS indicated one of the two peaksis the desired product. The DMF solution of reaction mixture wasdirectly injected into prep HPLC, only a portion of the product mixturewas purified.

Example 41

5-Dimethylaminomethylene-6,7-dihydro-5H-benzo[1,2,5]oxadiazol-4-one

A THF solution of 4,5,6,7-tetrahydro-2,1,3-benzoxadiazole-4-one 1 g(0.00725 mol) and Bredereck's reagent 1.89 g (0.0109 mol) was stirred atroom temperature for 2 days. The crystalline precipitate was filtered,washed with THF and dried, 0.8 g desired product was obtained, the yieldis 57%.

3-(4,5-dihydro-2-oxa-1,3,7,9-tetraaza-cyclopenta[α]naphthalen-8-ylamino)-phenol:The DMF solution of5-dimethylaminomethylene-6,7-dihydro-5H-benzo[1,2,5]oxadiazol-4-one 0.1g (0.00052 mol) and 3 equivalents of N-(3-hydroxy-phenyl)-guanidine wasstirred at 110° C. for overnight, the final product was purified bydirectly injecting the DMF reaction solution into prep HPLC.

Example 42

3-Dimethylamino-4-oxo-4,5,6,7-tetrahydro-benzo[c]thiophene-1-carbonitrile

A solution of3-methylsulfanyl-4-oxo-4,5,6,7-tetrahydro-benzo[c]thiophene-1-carbonitrile(0.446 g) and dimethylamine (excess) in THF were heated in a sealed tubeat 80° C. for 10 days. Aqueous work up and chromatographic purificationafforded 0.2 g of3-dimethylamino-4-oxo-4,5,6,7-tetrahydro-benzo[c]thiophene-1-carbonitrile(45%).

3-(3-Cyano-1-dimethylamino-4,5-dihydro-2-thia-7,9-diaza-cyclopenta-[α]naphthalen-8-ylamino)-benzenesulfonamide

-   1. A solution of    3-dimethylamino-4-oxo-4,5,6,7-tetrahydro-benzo[c]thiophene-1-carbonitrile    0.2 g (0.9 mmol) and excess of DMF-DMA in MeCN was refluxed    overnight. The solvent was evaporated and the crude material was    used for the next step without purification.-   2. The crude enaminone was heated together with    3-guanidino-benzenesulfonamide in DMF at 120° C. overnight.    Following aqueous work up, the product was purified by preparative    HPLC.

Example 43 Exemplary Synthesis of Compounds where X is —CH₂CH₂—, n is 0and Cy¹ is Optionally Substituted Phenyl

2-Dimethylaminomethylene-3,4-dihydro-2H-naphthalen-1-one (57)

A solution of 3,4-dihydro-2H-naphthalen-1-one in MeCN and 20 eq. ofDMF-DMA were stirred at 90° C. overnight. Thin layer chromatographyindicated there is almost no desired product, hence 1.2 equivalents ofBredereck's reagent was added, and the reaction mixture stirred at 80°C. overnight. The solvent was removed under reduced pressure and thecrude material was used for the next step with no further purification.

(5,6-Dihydro-benzo[h]quinazolin-2-yl)-phenyl-amine (58)

A solution of 2-dimethylaminomethylene-3,4-dihydro-2H-naphthalen-1-one100 mg in 5 ml DMF and 3 eq. of N-phenyl-guanidine was stirred at 110°C. for overnight, the product was purified by directly injecting the DMFreaction solution into prep HPLC.

BIOLOGICAL EXAMPLES Example 1 Aurora-2 Inhibition Assay

Compounds were screened for their ability to inhibit Aurora-2 using astandard coupled enzyme assay (Fox et al., Protein Sci., (1998) 7,2249). Assays were carried out in a mixture of 100 mM Hepes (pH7.5), 10mM MgCl₂, 1 mM DTT, 25 mM NaCl, 2.5 mM phosphoenolpyruvate, 300 μM NADH,30 μg/ml pyruvate kinase and 10 μg/ml lactate dehydrogenase. Finalsubstrate concentrations in the assay were 400 μM ATP (Sigma Chemicals)and 570 μM peptide (Kemptide, American Peptide, Sunnyvale, Calif.).Assays were carried out at 30° C. and in the presence of 40 nM Aurora-2.

An assay stock buffer solution was prepared containing all of thereagents listed above, with the exception of Aurora-2 and the testcompound of interest. 55 μl of the stock solution was placed in a 96well plate followed by addition of 2 μl of DMSO stock containing serialdilutions of the test compound (typically starting from a finalconcentration of 7.5 μM). The plate was preincubated for 10 minutes at30° C. and the reaction initiated by addition of 10 μl of Aurora-2.Initial reaction rates were determined with a Molecular DevicesSpectraMax Plus plate reader over a 10 minute time course. IC50 and Kidata were calculated from non-linear regression analysis using the Prismsoftware package (GraphPad Prism version 3.0cx for Macintosh, GraphPadSoftware, San Diego Calif., USA).

Compounds of the present invention were shown to inhibit Aurora-2 usingthe assay methods described above. In general, compounds of theinvention are effective for the inhibition of Aurora-2.

Example 2 CDK-2 Inhibition Assay

Compounds were screened for their ability to inhibit Cdk2/cyclin A usinga standard coupled enzyme assay (Fox et al., Protein Sci., (1998) 7,2249). Assays were carried out in a mixture of 25 mM Hepes (pH7.5), 10mM MgCl₂, 0.5 mM DTT, 2.5 mM phosphoenolpyruvate, 300 μM NADH, 30 μg/mlpyruvate kinase and 10 μg/ml lactate dehydrogenase. Final substrateconcentrations in the assay were 500 μM ATP (Sigma Chemicals) and 150 μMpeptide (Histone H1, Upstate Biotechnology, UK). Assays were carried outat 30° C. and in the presence of 9 nM Cdk2/cyclin A.

An assay stock buffer solution was prepared containing all of thereagents listed above, with the exception of ATP and the test compoundof interest. 60 μl of the stock solution was placed in a 96 well platefollowed by addition of 2 μl of DMSO stock containing serial dilutionsof the test compound (typically starting from a final concentration of7.5 μM). The plate was preincubated for 10 minutes at 30° C. and thereaction initiated by addition of 5 μl of ATP. Initial reaction rateswere determined with a Molecular Devices SpectraMax Plus plate readerover a 10 minute time course. IC50 and Ki data were calculated fromnon-linear regression analysis using the Prism software package(GraphPad Prism version 3.0cx for Macintosh, GraphPad Software, SanDiego Calif., USA).

Compounds of the present invention were shown to inhibit CDK-2 using theassay methods described above. In general, compounds of the inventionare effective for the inhibition of CDK-2.

Example 3 Inhibition of c-KIT

Compounds were screened for their ability to inhibit c-KIT activityusing a radiometric filter-binding assay. This assay monitors the ³³Pincorporation into a substrate poly(Glu, Tyr) 4:1 (pE4Y). Reactions werecarried out in a solution containing 100 mM HEPES (pH 7.5), 10 mM MgCl₂,25 mM NaCl, 1 mM DTT, 0.01% BSA and 2.5% DMSO. Final substrateconcentrations in the assay were 700 μM ATP and 0.5 mg/mL pE4Y (bothfrom Sigma Chemicals, St Louis, Mo.). The final concentration ofcompounds is generally between 0.01 and 5 μM. Typically, a 12-pointtitration was conducted by preparing serial dilutions from 10 mM DMSOstock of test compound. Reactions were carried out at room temperature.

Two assay solutions were prepared. Solution 1 contains 100 mM HEPES(pH7.5), 10 mM MgCl₂, 25 mM NaCl, 1 mg/ml pE4Y and 1.4 mM ATP(containing 0.5 μCi of [γ-³³P]ATP for each reaction). Solution 2contains 100 mM HEPES (pH7.5), 10 mM MgCl₂, 25 mM NaCl, 2 mM DTT, 0.02%BSA and 25 nM c-KIT. The assay was run on a 96 well plate by mixing 33μL of Solution 1 and 1.65 μL of the test compounds. The reaction wasinitiated with 33 μL of Solution 2. After incubation for 20 minutes atroom temperature, the reaction was stopped with 50 μL of 10% TCAcontaining 0.2 mM of ATP. All of the reaction volume was thentransferred to a filter plate and washed with 5% TCA by a Harvester9600from TOMTEC (Hamden, Conn.). The amount of ³³P incorporation into pE4ywas analyzed by a Packard TopCount Microplate Scintillation Counter(Meriden, Conn.). The data was fitted using Prism software to get anIC₅₀ or K_(i).

In general, compounds of the invention are effective for the inhibitionof c-KIT.

Example 4 cMET Inhibition Assay

Compounds were screened for their ability to inhibit cMet kinaseactivity using a standard coupled enzyme system (Fox et al., ProteinSci. 1998, 7, 2249). Reactions were carried out in a solution containing100 mM HEPES (pH 7.5), 10 mM MgCl₂, 25 mM NaCl, 300 μM NADH, 1 mM DTT,and 1.5% DMSO. Final substrate concentrations in the assay were 200 μMATP (Sigma Chemicals, St Louis, Mo.) and 10 μM polyGluTyr (SigmaChemical Company, St. Louis). Reactions were carried out at 30° C. and80 nM cMet. Final concentrations of the components of the coupled enzymesystem were 2.5 mM phosphoenolpyruvate, 300 μM NADH, 30 μg/ml pyruvatekinase and 10 μg/ml lactate dehydrogenase.

An assay stock buffer solution was prepared containing all of thereagents listed above with the exception of ATP and a test compound ofthe present invention. The assay stock buffer solution (175 μl) wasincubated in a 96 well plate with 5 μl of the test compound of thepresent invention at final concentrations spanning 0.006 μM to 12.5 μMat 30° C. for 10 min. Typically, a 12 point titration was conducted bypreparing serial dilutions (from 10 mM compound stocks) with DMSO of thetest compounds of the present invention in daughter plates. The reactionwas initiated by the addition of 20 μl of ATP (final concentration 200μM). Rates of reaction were obtained using a Molecular DevicesSpectramax plate reader (Sunnyvale, Calif.) over 10 min at 30° C. The K,values were determined from the rate data as a function of inhibitorconcentration.

Compounds of the present invention were shown to inhibit cMet using theassay methods described above. In general, compounds of the inventionare effective for the inhibition of cMet.

Example 5 ERK Inhibition Assay

Compounds were assayed for the inhibition of ERK2 by aspectrophotometric coupled-enzyme assay (Fox et al. Protein Sci. 1998,7, 2249). In this assay, a fixed concentration of activated ERK2 (10 nM)was incubated with various concentrations of a compound of the presentinvention in DMSO (2.5%) for 10 min. at 30° C. in 0.1 M HEPES buffer (pH7.5), containing 10 mM MgCl₂, 2.5 mM phosphoenolpyruvate, 200 μM NADH,150 μg/ml pyruvate kinase, 50 μg/ml lactate dehydrogenase, and 200 μMerktide peptide. The reaction was initiated by the addition of 65 μMATP. The rate of decrease of absorbance at 340 nM was monitored. The IC,values were determined from the rate data as a function of inhibitorconcentration.

In general, compounds of the invention are effective for the inhibitionof ERK-2.

Example 6 FLT-3 Inhibition Assay

Compounds were screened for their ability to inhibit FLT-3 activityusing a radiometric filter-binding assay. This assay monitors the ³³Pincorporation into a substrate poly (Glu, Tyr) 4:1 (pE4Y). Reactionswere carried out in a solution containing 100 mM HEPES (pH 7.5), 10 mMMgCl₂, 25 mM NaCl, 1 mM DTT, 0.01% BSA and 2.5% DMSO. Final substrateconcentrations in the assay were 90 μM ATP and 0.5 mg/ml pE4Y (both fromSigma Chemicals, St Louis, Mo.). The final concentration of a compoundof the present invention is generally between 0.01 and 5 μM. Typically,a 12-point titration was conducted by preparing serial dilutions from 10mM DMSO stock of test compound. Reactions were carried out at roomtemperature.

Two assay solutions were prepared. Solution 1 contains 100 mM HEPES (pH7.5), 10 mM MgCl₂, 25 mM NaCl, 1 mg/ml pE4Y and 180 μM ATP (containing0.3 μCi of [γ-³³P]ATP for each reaction). Solution 2 contains 100 mMHEPES (pH 7.5), 10 mM MgCl₂, 25 mM NaCl, 2 mM DTT, 0.02% BSA and 3 nMFLT-3. The assay was run on a 96 well plate by mixing 50 μl each ofSolution 1 and 2.5 ml of the compounds of the present invention. Thereaction was initiated with Solution 2. After incubation for 20 minutesat room temperature, the reaction was stopped with 50 μl of 20% TCAcontaining 0.4 mM of ATP. All of the reaction volume was thentransferred to a filter plate and washed with 5% TCA by a Harvester 9600from TOMTEC (Hamden, Conn.). The amount of ³³P incorporation into pE4ywas analyzed by a Packard Top Count Microplate Scintillation Counter(Meriden, Conn.). The data was fitted using Prism software to get anIC₅₀ or K_(i).

Compounds of the present invention were shown to inhibit FLT-3 using theassay methods described above. In general, compounds of the inventionare effective for the inhibition of FLT-3.

Example 7 GSK-3 Inhibition Assay

Compounds were screened for their ability to inhibit GSK-3β using astandard coupled enzyme assay (Fox et al., Protein Sci., (1998) 7,2249). Assays were carried out in a mixture of 100 mM Hepes (pH7.5), 10mM MgCl₂, 1 mM DTT, 25 mM NaCl, 2.5 mM phosphoenolpyruvate, 300 μM NADH,30 μg/ml pyruvate kinase and 10 μg/ml lactate dehydrogenase. Finalsubstrate concentrations in the assay were 60 μM ATP (Sigma Chemicals)and 300 μM peptide (HSSPHQS(PO₃H₂)EDEEE, American Peptide, Sunnyvale,Calif.). Assays were carried out at 30° C. and in the presence of 35 nMGSK-3β.

An assay stock buffer solution was prepared containing all of thereagents listed above, with the exception of ATP and the test compoundof interest. 60 μl of the stock solution was placed in a 96 well platefollowed by addition of 2 μl of DMSO stock containing serial dilutionsof the test compound (typically starting from a final concentration of7.5 μM). The plate was preincubated for 10 minutes at 30° C. and thereaction initiated by addition of 5 μl of ATP. Initial reaction rateswere determined with a Molecular Devices SpectraMax Plus plate readerover a 10 minute time course. IC50 and Ki data were calculated fromnon-linear regression analysis using the Prism software package(GraphPad Prism version 3.0cx for Macintosh, GraphPad Software, SanDiego Calif., USA).

In general, compounds of the invention are effective for the inhibitionof GSK-3.

Example 8 JAK Inhibition Assay

Method A

Compounds of the present invention were screened for their ability toinhibit JAK activity using the method described by G. R. Brown et al.,Bioorg. Med. Chem. Lett. 2000, 10, 575-579 in the following manner. IntoMaxisorb plates, previously coated at 4° C. with Poly (Glu, Ala, Tyr)6:3:1 then washed with phosphate buffered saline 0.05% and Tween (PBST),was added 2 μM ATP, 5 mM MgCl₂, and a solution of a compound of thepresent invention in DMSO. The reaction was started with JAK enzyme andthe plates incubated for 60 minutes at 30° C. The plates were thenwashed with PBST, 100 μl HRP-Conjugated 4G10 antibody was added, and theplate incubated for 90 minutes at 30° C. The plate was again washed withPBST, 100 μl TMB solution was added, and the plates were incubated foranother 30 minutes at 30° C. Sulfuric acid (100 μl of a 1M solution) wasadded to stop the reaction and the plate was read at 450 nm to obtainthe optical densities for analysis to determine IC₅₀ values and K_(i)values.

Method B

JAK-3 Assay Components:

-   “kinase buffer”: 100 mM HEPES pH 7.4; 1 mM DTT; 10 mM MgCl₂; 25 mM    NaCl; 0.01%; BSA.-   1 nM JAK3 (enzyme)-   1 uM poly(Glu)₄Tyr (substrate)-   5 uM ATP (substrate, 200 uCi/umole ATP)    Procedure:

To each well of a 96 well polycarbonate plate is added 1.5 ul of acandidate JAK3 inhibitor along with 50 ul of kinase buffer containing 2uM poly(Glu)₄Tyr and 10 uM ATP. This is then mixed and 50 ul of kinasebuffer containing 2 nM JAK-3 enzyme is added to start the reaction.After 20 minutes at room temperature (25 C), the reaction is stoppedwith 50 ul of 20% trichloroacetic acid (TCA) that also contains 0.4 mMATP. The entire content of each well is then transferred to a 96 wellglass fiber filter plate using a TomTek Cell Harvester. After washing,60 ul of scintillation fluid is added and ³³P incorporation detected ona Perkin Elmer TopCount.

JAK-2 Assay:

As above except that final poly(Glu)₄Tyr concentration is 15 uM andfinal ATP concentration is 12 uM.

In general, compounds of the invention are effective for the inhibitionof JAK (particularly JAK-3).

Example 9 SRC Inhibition Assay

Compounds were screened for their ability to inhibit Src using astandard coupled enzyme assay (Fox et al., Protein Sci., (1998) 7,2249). Assays were carried out in a mixture of 25 mM Hepes (pH7.5), 10mM MgCl₂, 2.2 mM DTT, 2.5 mM phosphoenolpyruvate, 300 μM NADH, 30 μg/mlpyruvate kinase and 10 μg/ml lactate dehydrogenase. Final substrateconcentrations in the assay were 100 μM ATP (Sigma Chemicals) and 0.28mg/ml peptide (poly 4Glu:Tyr, Sigma Chemicals). Assays were carried outat 30° C. and in the presence of 25 nM Src.

An assay stock buffer solution was prepared containing all of thereagents listed above, with the exception of peptide and the testcompound of interest. 60 μl of the stock solution was placed in a 96well plate followed by addition of 2 μl of DMSO stock containing serialdilutions of the test compound (typically starting from a finalconcentration of 7.5 μM). The plate was preincubated for 10 minutes at30° C. and the reaction initiated by addition of 5 μl of peptide.Initial reaction rates were determined with a Molecular DevicesSpectraMax Plus plate reader over a 10 minute time course. IC50 and Kidata were calculated from non-linear regression analysis using the Prismsoftware package (GraphPad Prism version 3.0cx for Macintosh, GraphPadSoftware, San Diego Calif., USA).

In general, compounds of the invention are effective for the inhibitionof SRC.

Example 10 LCK Inhibition Assay

The compounds were assayed as inhibitors of lck kinase purified frombovine thymus (from Upstate Biotechnology, cat. no. 14-106). Lck kinaseactivity was monitored by following the incorporation of ³³P from ATPinto the tyrosine of a random poly Glu-Tyr polymer substrate ofcomposition, Glu:Tyr=4:1 (Sigma, cat. no. P-0275). The following werethe final concentrations of the assay components: 0.05 M HEPES, pH 7.6,10 mM MgCl₂, 2 mM DTT, 0.25 mg/ml BSA, 10 μM ATP (1-2 μCi ³³P-ATP perreaction), 5 mg/ml poly Glu-Tyr, and 1-2 units of lck kinase. In atypical assay, all the reaction components with the exception of ATPwere pre-mixed and aliquoted into assay plate wells. Inhibitorsdissolved in DMSO were added to the wells to give a final DMSOconcentration of 2.5%. The assay plate was incubated at 30° C. for 10 mMbefore initiating the reaction with ³³P-ATP. After 20 mM of reaction,the reactions were quenched with 150 μl of 10% trichloroacetic acid(TCA) containing 20 mM Na₃PO4. The quenched samples were thentransferred to a 96-well filter plate (Whatman, UNI-Filter GF/F GlassFiber Filter, cat no. 7700-3310) installed on a filter plate vacuummanifold. Filter plates were washed four times with 10% TCA containing20 mM Na₃PO₄ and then 4 times with methanol. 2000 of scintillation fluidwas then added to each well. The plates were sealed and the amount ofradioactivity associated with the filters was quantified on a TopCountscintillation counter.

In general, compounds of the invention are effective for the inhibitionof LCK.

Example 11 SYK Inhibition Assay

Compounds were screened for their ability to inhibit Syk using astandard coupled enzyme assay (Fox et al., Protein Sci., (1998) 7,2249). Assays were carried out in a mixture of 100 mM Hepes (pH7.5), 10mM MgCl₂, 2 mM DTT, 25 mM NaCl, 2.5 mM phosphoenolpyruvate, 300 μM NADH,30 μg/ml pyruvate kinase and 10 μg/ml lactate dehydrogenase. Finalsubstrate concentrations in the assay were 100 μM ATP (Sigma Chemicals)and 20 nM peptide (poly 4Glu:Tyr, Sigma Chemicals). Assays were carriedout at 30° C. and in the presence of 20 nM Syk.

An assay stock buffer solution was prepared containing all of thereagents listed above, with the exception of Syk enzyme and the testcompound of interest. 55 μl of the stock solution was placed in a 96well plate followed by addition of 2 μl of DMSO stock containing serialdilutions of the test compound (typically starting from a finalconcentration of 7.5 μM). The plate was preincubated for 10 minutes at30° C. and the reaction initiated by addition of 10 μl of Syk enzyme.Initial reaction rates were determined with a Molecular DevicesSpectraMax Plus plate reader over a 10 minute time course. IC50 and Kidata were calculated from non-linear regression analysis using the Prismsoftware package (GraphPad Prism version 3.0cx for Macintosh, GraphPadSoftware, San Diego Calif., USA).

In general, compounds of the invention are effective for the inhibitionof SYK.

Example 12 ITK Inhibition Assay

The compounds of the present invention were evaluated as inhibitors ofhuman Itk kinase using either a radioactivity-based, spectrophotometricor alphascreen assay.

Itk Inhibition Assay: Radioactivity-Based Assay (Method A)

Assays were carried out in a mixture of 100 mM HEPES (pH 7.5), 10 mMMgCl₂, 25 mM NaCl, 0.01% BSA and 1 mM DTT. Final substrateconcentrations were 15 μM [γ-³³P]ATP (400 μCi ³³P ATP/μmol ATP, AmershamPharmacia Biotech/Sigma Chemicals) and 2 μM peptide (SAM68 proteinΔ332-443). Assays were carried out at 25° C. in the presence of 30 nMItk. An assay stock buffer solution was prepared containing all of thereagents listed above, with the exception of ATP and the test compoundof interest. 50 μL of the stock solution was placed in a 96 well platefollowed by addition of 1.5 μL of DMSO stock containing serial dilutionsof the test compound (typically starting from a final concentration of15 μM with 2-fold serial dilutions) in duplicate (final DMSOconcentration 1.5%). The plate was pre-incubated for 10 minutes at 25°C. and the reaction initiated by addition of 50 μL [γ-³³P]ATP (finalconcentration 15 μM).

The reaction was stopped after 10 minutes by the addition of 50 μL of aTCA/ATP mixture (20% TCA, 0.4 mM ATP). A Unifilter GF/C 96 well plate(Perkin Elmer Life Sciences, Cat no. 6005174) was pretreated with 50 μLMilli Q water prior to the addition of the entire reaction mixture (150μL). The plate was washed with 2004 Milli Q water followed by 2004 of aTCA/ATP mixture (5% TCA, 1 mM ATP). This wash cycle was repeated afurther 2 times. After drying, 30 μL Optiphase ‘SuperMix’ liquidscintillation cocktail (Perkin Elmer) was added to the well prior toscintillation counting (1450 Microbeta Liquid Scintillation Counter,Wallac).

IC50 data were calculated from non-linear regression analysis of theinitial rate data using the Prism software package (GraphPad Prismversion 3.0cx for Macintosh, GraphPad Software, San Diego Calif., USA).

Itk Inhibition Assay: Alphascreen Assay (Method B)

Assays were carried out in a mixture of 20 mM MOPS (pH 7.0), 10 mMMgCl₂, 0.1% BSA and 1 mM DTT. Final substrate concentrations in theassay were 100 μM ATP (Sigma Chemicals) and 2 μM peptide (BiotinylatedSAM68 Δ332-443). Assays were carried out at 25° C. and in the presenceof 10 nM Itk. An assay stock buffer solution was prepared containing allof the reagents listed above, with the exception of ATP and the testcompound of interest. 25 μL of the stock solution was placed in eachwell of a 96 well plate followed by 1 μL of DMSO containing serialdilutions of the test compound (typically starting from a finalconcentration of 15 μM) in duplicate (final DMSO concentration 2%). Theplate was preincubated for 10 minutes at 25° C. and the reactioninitiated by addition of 25 μL ATP (final concentration 100 μM).Background counts were determined by the addition of 5 μL 500 mM EDTA tocontrol wells containing assay stock buffer and DMSO prior to initiationwith ATP. The reaction was stopped after 30 minutes by diluting thereaction 225-fold into MOPS buffer (20 mM MOPS (pH 7.0), 1 mM DTT, 10 mMMgCl₂, 0.1% BSA) containing 50 mM EDTA to bring the final concentrationof peptide to 9 nM.

AlphaScreen™ reagents were prepared according to the manufacturersinstructions (AlphaScreen™ phosphotyrosine (P-Tyr-100) assay kit,PerkinElmer catalogue number 6760620C). Under subdued lighting, 20 μL ofAlphaScreen™ reagents were placed in each well of a white half area 96well plate (Corning Inc.—COSTAR 3693) with 30 μL of the stopped, dilutedkinase reactions. Plates were incubated in the dark for 60 minutes priorto reading on a Fusion Alpha plate reader (PerkinElmer).

After removing mean background values for all of the data points,Ki(app) data were calculated from non-linear regression analysis usingthe Prism software package (GraphPad Prism version 3.0cx for Macintosh,GraphPad Software, San Diego Calif., USA).

Itk Inhibition Assay: Spectrophotometric Assay (Method C)

Compounds were screened for their ability to inhibit Itk using astandard coupled enzyme assay (Fox et al., Protein Sci., (1998) 7,2249).

Assays were carried out in a mixture of 20 mM MOPS (pH 7.0), 10 mMMgCl₂, 0.1% BSA, 1 mM DTT, 2.5 mM phosphoenolpyruvate, 300 μM NADH, 30μg/ml pyruvate kinase and 10 μg/ml lactate dehydrogenase. Finalsubstrate concentrations in the assay were 100 μM ATP (Sigma Chemicals)and 3 μM peptide (Biotinylated SAM68 Δ332-443). Assays were carried outat 25° C. and in the presence of 100 nM Itk.

An assay stock buffer solution was prepared containing all of thereagents listed above, with the exception of ATP and the test compoundof interest. 60 μl of the stock solution was placed in a 96 well platefollowed by addition of 2 μl of DMSO stock containing serial dilutionsof the test compound (typically starting from a final concentration of15 μM). The plate was preincubated for 10 minutes at 25° C. and thereaction initiated by addition of 5 μl of ATP. Initial reaction rateswere determined with a Molecular Devices SpectraMax Plus plate readerover a 10 minute time course. IC50 and Ki data were calculated fromnon-linear regression analysis using the Prism software package(GraphPad Prism version 3.0cx for Macintosh, GraphPad Software, SanDiego Calif., USA).

In general, compounds of the invention are effective for the inhibitionof Itk. Preferred compounds showed IC50 below 1 μM in the radioactiveincorporation assay (Method A) (I-1, I-2, I-3, I-4, I-5, I-6, I-7, I-8,I-9, I-10, I-11, I-12, I-13, I-14, I-15, I-17, I-18, I-19, I-20, I-21,I-23, I-24, I-25, I-27, I-28, I-29. Preferred compounds showed Ki below1 μM in the AlphaScreen™ assay (Method B) (I-36, I-41, II-13, II-34,II-35, II-36, II-37, II-38, II-39, III-4, III-31, III-32, III-33,III-34, III-35, III-38, III-39, III-40, IV-4, V-4, V-36, V-37, V-38,V-39, V-40, V-41, V-42, V-43, V-44, V-45, V-46, V-47, V-48, V-49, V-50,V-51, V-52, V-54). Preferred compounds showed Ki below 1 μM in thestandard coupled enzyme assay (Method C) (V-55, V-56, V-58, V-59, V-60,V-61, V-63, V-67, V-68, V-69, V-70, V-71, V-72, V-73, V-74, V-76, V-77,V-78, V-79, V-80, V-81, V-82, V-83, V-84, V-85, V-86, V-87, V-88, V-89,V-90, V-91, V-92, V-93, V-94, V-95, V-96, V-97, V-98, V-100, V-101,V-102, V-103, V-104, V-105, V-106, V-107, V-108, V-109, V-110, V-111,V-112, V-113, V-114, V-115, V-116, V-117, V-118, V-119, V-120, V-121,V-122, V-125, V-127, V-129, V-130, V-131, V-132, V-133, V-134, V-135,V-136, V-137, V-138, V-139, V-140, V-141, V-142, V-143, V-144, V-145,V-146, V-147, V-148, V-149, V-150, V-151, V-152, V-153, V-154, V-155,V-156, V-157, V-159, V-162, V-165, V-166, V-167, V-170, V-172, V-173,V-175, V-176, V-177, V-178, V-179, V-180, V-182).

Example 13 BTK Inhibition Assay

The compounds of the present invention were evaluated as inhibitors ofhuman Btk kinase using either a radioactivity-based or alphascreenassay.

Btk Inhibition Assay: Radioactivity-Based Assay

Assays were carried out in a mixture of 20 mM MOPS (pH 7.0), 10 mMMgCl₂, 0.1% BSA and 1 mM DTT. Final substrate concentrations in theassay were 50 μM [γ-³³P]ATP (200 μCi ³³P ATP/μmol ATP, AmershamPharmacia Biotech, Amersham, UK/Sigma Chemicals) and 2 μM peptide (SAM68Δ332-443). Assays were carried out at 25° C. and in the presence of 25nM Btk. An assay stock buffer solution was prepared containing all ofthe reagents listed above, with the exception of the peptide and thetest compound of interest. 75 μL of the stock solution was placed in a96 well plate followed by addition of 2 μL of DMSO stock containingserial dilutions of the test compound (typically starting from a finalconcentration of 15 μM) in duplicate (final DMSO concentration 2%). Theplate was preincubated for 15 minutes at 25° C. and the reactioninitiated by addition of 25 μL peptide (final concentration 2 μM).Background counts were determined by the addition of 100 μL 0.2Mphosphoric acid+0.01% TWEEN to control wells containing assay stockbuffer and DMSO prior to initiation with peptide.

The reaction was stopped after 10 minutes by the addition of 100 μL 0.2Mphosphoric acid+0.01% TWEEN. A multiscreen phosphocellulose filter96-well plate (Millipore, Cat no. MAPHN0B50) was pretreated with 100 μL0.2M phosphoric acid+0.01% TWEEN 20 prior to the addition of 170 μL ofthe stopped assay mixture. The plate was washed with 4×200 μL 0.2Mphosphoric acid+0.01% TWEEN 20. After drying, 30 μL Optiphase ‘SuperMix’liquid scintillation cocktail (Perkin Elmer) was added to the well priorto scintillation counting (1450 Microbeta Liquid Scintillation Counter,Wallac).

After removing mean background values for all of the data points,Ki(app) data were calculated from non-linear regression analysis usingthe Prism software package (GraphPad Prism version 3.0cx for Macintosh,GraphPad Software, San Diego Calif., USA).

Btk Inhibition Assay: Alphascreen Assay

Assays were carried out in a mixture of 20 mM MOPS (pH 7.0), 10 mMMgCl₂, 0.1% BSA and 1 mM DTT. Final substrate concentrations in theassay were 50 μM ATP (Sigma Chemicals) and 2 μM peptide (BiotinylatedSAM68 Δ332-443). Assays were carried out at 25° C. and in the presenceof 25 nM Btk. An assay stock buffer solution was prepared containing allof the reagents listed above, with the exception of peptide and the testcompound of interest. 37.5 μL, of the stock solution was placed in eachwell of a 96 well plate followed by 1 μL of DMSO containing serialdilutions of the test compound (typically starting from a finalconcentration of 15 μM) in duplicate (final DMSO concentration 2%). Theplate was preincubated for 15 minutes at 25° C. and the reactioninitiated by addition of 12.5 μL peptide (final concentration 2 μM).Background counts were determined by the addition of 5 μL 500 mM EDTA tocontrol wells containing assay stock buffer and DMSO prior to initiationwith Biotin-SAM68.

The reaction was stopped after 30 minutes by diluting the reaction225-fold into MOPS buffer (20 mM MOPS (pH 7.0), 1 mM DTT, 10 mM MgCl₂,0.1% BSA) containing 50 mM EDTA to bring the final concentration ofpeptide to 9 nM.

AlphaScreen™ reagents were prepared according to the manufacturersinstructions (AlphaScreen™ phosphotyrosine (P-Tyr-100) assay kit,PerkinElmer catalogue number 6760620C). Under subdued lighting, 20 μL ofAlphaScreen™ reagents were placed in each well of a white half area 96well plate (Corning Inc.—COSTAR 3693) with 30 μL of the stopped, dilutedkinase reactions. Plates were incubated in the dark for 60 minutes priorto reading on a Fusion Alpha plate reader (PerkinElmer).

After removing mean background values for all of the data points,Ki(app) data were calculated from non-linear regression analysis usingthe Prism software package (GraphPad Prism version 3.0cx for Macintosh,GraphPad Software, San Diego Calif., USA).

In general, compounds of the invention are effective for the inhibitionof Btk.

While we have described a number of embodiments of this invention, it isapparent that our basic examples may be altered to provide otherembodiments which utilize the compounds and methods of this invention.Therefore, it will be appreciated that the scope of this invention is tobe defined by the appended claims rather than by the specificembodiments that have been represented by way of example above.

1. A compound of formula I:

or a pharmaceutically acceptable salt thereof, wherein: X is —(CH₂)₂—,wherein one or more of the hydrogen substituents are optionally andindependently replaced with m occurrences of —WR^(y), wherein m is aninteger selected from 0 to 4; R¹ and R², taken together with the carbonatoms to which they are bound form an optionally substituted 5-memberedheteroaryl thiophene ring, wherein the ring formed by R¹ and R² takentogether is optionally and independently substituted at one or morecarbon atoms with up to 2 occurrences of —WR^(y); each occurrence of Wis independently a bond or is a C₁-C₆ alkylidene chain wherein up to twonon-adjacent methylene units of W are independently optionally replacedby CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO, NRCO₂, NRCONR, SO,SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR; and each occurrence of R^(y) isindependently selected from R′, halogen, NO₂, or CN, or —WR^(y) is ═O,═S, or ═NR′; T is CHR′, CH₂CH(R′), —S(═O)₂, or C(═O); n is 0 or 1; Cy¹is a 3-7 membered saturated, partially unsaturated, or fully unsaturatedmonocyclic ring having 0-3 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, or an 8-10 membered saturated, partiallyunsaturated, or fully unsaturated bicyclic ring system having 0-5heteroatoms independently selected from nitrogen, oxygen, or sulfur,wherein Cy¹ is optionally and independently substituted at one or morecarbon atoms with k independent occurrences of -QR^(x) wherein k is aninteger selected from 0 to 5; and at one or more substitutable nitrogenatoms with —R⁴; wherein Q is a bond or is a C₁-C₆ alkylidene chainwherein up to two non-adjacent methylene units of Q are independentlyoptionally replaced by CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO,NRCO₂, NRCONR, SO, SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR; and eachoccurrence of R^(x) is independently selected from R′, halogen, NO₂,CF₃, or CN, or -QR^(x) is ═O, ═S, or ═NR′; each occurrence of R³ and R⁴is independently R′, —COR′, —CO₂(C₁₋₆ aliphatic), —CON(R′)₂, or —SO₂R′;each occurrence of R is independently selected from hydrogen or anoptionally substituted C₁₋₆ aliphatic group; and each occurrence of R′is independently selected from hydrogen or an optionally substitutedgroup selected from C₁₋₈ aliphatic, C₆₋₁₀ aryl, a heteroaryl ring having5-10 ring atoms, or a heterocyclyl ring having 3-10 ring atoms, orwherein R and R′ taken together with the atom(s) to which they arebound, or two occurrences of R′ taken together with the atom(s) to whichthey are bound, form an optionally substituted 3-8 membered cycloalkyl,heterocyclyl, aryl, or heteroaryl ring having 0-3 heteroatomsindependently selected from nitrogen, oxygen, or sulfur; and the dashedbond represents a single or double bond, as valency permits; providedthat: when n is 0, Cy¹ is 3, 4, 5-trimethoxyphenyl, and X is —(CH₂)₂—,then R¹ and R², taken together with the carbon atoms to which they arebound, do not form an unsubstituted thieno ring.
 2. A compound offormula I:

or a pharmaceutically acceptable salt thereof, wherein: X is —(CH₂)₂—,wherein one or more of the hydrogen substituents are optionally andindependently replaced with m occurrences of —WR^(y), wherein m is aninteger selected from 0 to 4; R¹ and R², taken together with the carbonatoms to which they are bound form an optionally substituted 5-memberedheteroaryl thiophene ring, wherein the ring formed by R¹ and R² takentogether is optionally and independently substituted at one or morecarbon atoms with up to 2 occurrences of —WR^(y); each occurrence of Wis independently a bond or is a C₁-C₆ alkylidene chain wherein up to twonon-adjacent methylene units of W are independently optionally replacedby CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO, NRCO₂, NRCONR, SO,SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR; and each occurrence of R^(y) isindependently selected from R′, halogen, NO₂, or CN, or —WR^(y) is ═O,═S, or ═NR′; T is CHR′, CH₂CH(R′), —S(═O)₂, C(═O), CHR′CH₂; CH₂CH₂CH₂,or CH₂CHR′CH₂; n is 0 or 1; Cy¹ is a 3-7 membered saturated, partiallyunsaturated, or fully unsaturated monocyclic ring having 0-3 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or an 8-10membered saturated, partially unsaturated, or fully unsaturated bicyclicring system having 0-5 heteroatoms independently selected from nitrogen,oxygen, or sulfur, wherein Cy¹ is optionally and independentlysubstituted at one or more carbon atoms with k independent occurrencesof -QR^(x); wherein k is an integer selected from 0 to 5; and at one ormore substitutable nitrogen atoms with —R⁴; wherein Q is a bond or is aC₁-C₆ alkylidene chain wherein up to two non-adjacent methylene units ofQ arc independently optionally replaced by CO, CO₂, COCO, CONR, OCONR,NRNR, NRNRCO, NRCO, NRCO₂, NRCONR, SO, SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S,or NR; and each occurrence of R^(x) is independently selected from R′,halogen, NO₂, CF₃, or CN, or -QR^(x) is ═O, ═S, or ═NR′; each occurrenceof R³ and R⁴ is independently R′, —COR′, —CO₂(C₁₋₆ aliphatic),—CON(R′)₂, or —SO₂R′; each occurrence of R is independently selectedfrom hydrogen or an optionally substituted C₁₋₆ aliphatic group; andeach occurrence of R′ is independently selected from hydrogen or anoptionally substituted group selected from C₁₋₈ aliphatic, C₆₋₁₀ aryl, aheteroaryl ring having 5-10 ring atoms, or a heterocyclyl ring having3-10 ring atoms, or wherein R and R′ taken together with the atom(s) towhich they are bound, or two occurrences of R′ taken together with theatom(s) to which they are bound, form an optionally substituted 3-8membered cycloalkyl, heterocyclyl, aryl, or heteroaryl ring having 0-3heteroatoms independently selected from nitrogen, oxygen, or sulfur; andthe dashed bond represents a single or double bond, as valency permits;provided that: when n is 0 and Cy¹ is 3,4,5-trimethoxyphenyl, X is—(CH₂)₂—, then R¹ and R², taken together with the carbon atoms to whichthey are bound, do not form an unsubstituted thieno ring.
 3. Thecompound of claim 1 or claim 2, wherein: when n is 0, Cy¹ is anoptionally substituted phenyl, benzotriazolyl, benzothiazolyl, indolyl,or pyridyl group, and X is —(CH₂)₂—, optionally substituted with one ormore halogen atoms or C₁₋₃alkyl groups, then R¹ and R², taken togetherwith the carbon atoms to which they are bound do not form a thienylgroup that is unsubstituted or is substituted with a substituent otherthan a 3-7 membered saturated, partially unsaturated, or fullyunsaturated monocyclic ring having 0-3 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur, or an 8-10 memberedsaturated, partially unsaturated, or fully unsaturated bicyclic ringsystem having 0-5 heteroatoms independently selected from nitrogen,oxygen, or sulfur.
 4. The compound of claim 1 or claim 2, wherein R¹ andR², taken together with the carbon atoms to which they are bound form anoptionally substituted 5-membered heteroaryl ring selected from iv-1,iv-2, and xviii:


5. The compound of claim 1 or claim 2, wherein each occurrence ofWR^(y), when present, is independently halogen, CN, NO₂, —N(R′)₂,—-CH₂N(R′)₂, —OR′, —CH₂OR′, —SR′, —CH₂SR′, —COOR′, —NRCOR′, —CON(R′)₂,or —S(O)₂N(R′)₂, or an optionally substituted group selected fromC₁₋₄alkyl, aryl, heteroaryl.
 6. The compound of claim 1 or claim 2,wherein —WR^(y) groups are each independently F, Cl, Br, I, Me, Et, —CN,—OMe, —SMe, —NMe₂, —NEt₂, —COOMe, —COOH, —OH, —SO₂NH₂, —CON(CH₃)₂,—CO(optionally substituted N-piperazinyl), —CO(N-morpholinyl),—CO(N-piperidinyl), —CH₂N(Me)₂, —CH₂N(Et)₂, or an optionally substitutedgroup selected from C₁₋₄alkoxy, phenyl, phenyloxy, benzyl, benzyloxy,pyridyl, pyrimidinyl, thiophene, N-morpholinyl, N-piperidinyl,N-piperazinyl, or furanyl.
 7. The compound of claim 1 or claim 2,wherein —WR^(y) groups are each independently CF₃, F, Cl, Br, I, Me, Et,—CN, —OMe, —SMc, —NMe₂, —NEt₂, —COOMe, —COOH, —OH, —SO₂NH₂, —CON(CH₃)₂,—CO(optionally substituted N-piperazinyl, —CO(N-morpholinyl),—CO(N-piperidinyl), —CO(pyrrolidinyl), —CO(N(H)pyrrolidinyl),—CH₂N(Me)₂, —CH₂N(Et)₂, or an optionally substituted group selected fromC₁₋₄alkoxy, phenyl, phenyloxy, benzyl, benzyloxy, pyridyl, pyrimidinyl,thiophene, N-morpholinyl, N-piperidinyl, N-piperazinyl, furanyl,pyrrolidinyl, or N(H)pyrrolidinyl.
 8. The compound of claim 1 or claim2, wherein m is 1 and WR^(y) is an optionally substituted aryl orheteroaryl group, wherein substituents for the optionally substitutedaryl or heteroaryl group are independently selected from —VR^(v);wherein V is a bond or is a C₁-C₆ alkylidene chain wherein up to twonon-adjacent methylene units of V are independently optionally replacedby CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO, NRCO₂, NRCONR, SO,SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR; and each occurrence of R^(v) isindependently selected from R′, halogen, NO₂, or CN, or -QR^(x) is ═O,═S, or ═NR′.
 9. The compound of claim 1 or claim 2, wherein m is 1 andWR^(y) is —CH₂N(R′)₂, —N(R′)₂, or —CON(R′)₂.
 10. The compound of claim 1or claim 2, wherein T groups, when present are —CH₂—, —CH₂CH₂, —CO—, or—SO₂—.
 11. The compound of claim 1 or claim 2, wherein T groups, whenpresent are —C(H)(CH₃)—, —C(H)(CH₃)CH₂—, —CH₂C(H)(CH₃)—, or —CH₂CH₂CH₂—.12. The compound of claim 1 or claim 2, wherein n is 0 and T is absent.13. The compound of claim 1 or claim 2, wherein T is absent (n=0) or Tis —CO—.
 14. The compound of claim 1 or claim 2, wherein Cy¹ is selectedfrom one of the following groups:


15. The compound of claim 1 or claim 2, wherein Cy¹ is selected from:


16. The compound of claim 1 or claim 2, wherein Cy¹ is selected from:


17. The compound of claim 1 or claim 2, wherein Cy¹ is phenyl (a), andcompounds have the formula I-A:


18. The compound of claim 1 or claim 2, wherein QR^(X) groups, whenpresent, are each independently halogen, CN, NO₂, or an optionallysubstituted group selected from C₁₋₄alkyl, aryl, aralkyl, —N(R′)₂,—CH₂N(R′)₂, —OR′, —CH₂OR′, —SR′, —CH₂SR′, —COOR′, —NRCOR′, —CON(R′)₂,—SO₂R′, NR′SO₂R′, or —SO₂N(R′)₂.
 19. The compound of claim 1 or claim 2,wherein QR^(X) groups, when present, are each independently halogen, CN,NO₂, or an optionally substituted group selected from C₁₋₄alkyl, —CF₃,aryl, heteroaryl, aralkyl, —N(R′)₂, —CH₂N(R′)₂, —OR′, —CH₂OR′, —SR′,—CH₂SR′, —COOR′, —NRCOR′, —CON(R′)₂, —SO₂R′, NR′SO₂R′, SO₂N═R′, or—SO₂N(R′)₂.
 20. The compound of claim 1 or claim 2, wherein QR^(X)groups are each independently Cl, Br, F, CF₃, methyl, ethyl, isopropyl,n-propyl, n-butyl, tert-butyl, NO₂, —OH, —SO₂NH₂, SO₂CH₃, NH₂, SO₂NHCH₃,NHSO₂CH₃, or an optionally substituted group selected from C₁₋₄alkoxy,phenyl, phenyloxy, benzyl, or benzyloxy.
 21. The compound of claim 1 orclaim 2, wherein X is —(CH₂)₂—, and the compounds have the structure ofIII:


22. The compound of claim 21 wherein Cy¹ is optionally substitutedphenyl, and compounds have the formula III-A:


23. The compound of claim 21, wherein R¹ and R², taken together with thecarbon atoms to which they are bound form an optionally substituted5-membered heteroaryl ring selected from thiophenyl.
 24. The compound ofclaim 21, wherein R¹ and R², taken together with the carbon atoms towhich they are bound form an optionally substituted 5-memberedheteroaryl ring selected from thiophenyl and compounds have one of theformulae III-G, III-H, or III-I:


25. The compound of claim 21, wherein Cy¹ is phenyl, optionallysubstituted with 0-3 occurrences of QR^(X).
 26. The compound of claim21, wherein n is 0, or n is 1 and T is CH₂, —CH₂CH₂—, —CO— or —SO₂—; kis 0-3; and each occurrence of QR^(X) is independently halogen, CN, NO₂,or an optionally substituted group selected from C₁₋₄alkyl, aryl,aralkyl, —N(R′)₂, —CH₂N(R′)₂, —OR′, —CH₂OR′, —SR′, —CH₂SR′, —COOR′,—NRCOR′, —CON(R′)₂, —SO₂R′, NR′SO₂R′, or —SO₂N(R′)₂.
 27. The compound ofclaim 21, wherein m is 0-3; and each occurrence of WR^(y) isindependently halogen, CN, NO₂, or an optionally substituted groupselected from C₁₋₄alkyl, aryl, heteroaryl, —N(R′)₂, —CH₂N(R′)₂, —OR′,—CH₂OR′, —SR′, —CH₂SR′, —COOR′, —NRCOR′, —CON(R′)₂, or —S(O)₂N(R′)₂. 28.The compound of claim 27, wherein —WR^(y) groups are each independentlyF, Cl, Br, I, Me, Et, —CN, —OMe, —SMe, —NMe₂, —NEt₂, —COOMe, —COOH, —OH,—SO₂NH₂, —CON(CH₃)₂, —CO(optionally substituted N-piperazinyl),—CO(N-morpholinyl), —CO(N-piperidinyl), —CH₂N(Me)², —CH₂N(Et)₂, or anoptionally substituted group selected from C₁₋₄alkoxy, phenyl,phcnyloxy, benzyl, benzyloxy, pyridyl, pyrimidinyl, thiophene,N-morpholinyl, N-piperidinyl, N-piperazinyl, or furanyl.
 29. Thecompound claim 27, wherein —WR^(y) groups arc each independently CF₃, F,Cl, Br, I, Me, Et, —CN, —OMc, —SMe, —NMe₂, —NEt₂, —COOMe, —COOH, —OH,—SO₂NH₂, —CON(CH₃)₂, —CO(optionally substituted N-piperazinyl),—CO(N-morpholinyl), —CO(N-piperidinyl), —CO(pyrrolidinyl),—CO(N(H)pyrrolidinyl), —CH₂N(Me)₂, —CH₂N(Et)₂, or an optionallysubstituted group selected from C₁₋₄alkoxy, phenyl, phenyloxy, benzyl,benzyloxy, pyridyl, pyrimidinyl, thiophene, N-morpholinyl,N-piperidinyl, N-piperazinyl, furanyl, pyrrolidinyl, orN(H)pyrrolidinyl.
 30. The compound of claim 27, wherein one occurrenceof WR^(y) is an optionally substituted aryl or heteroaryl group, denotedby Ar¹ in formula III-H-i below, wherein m is 1-3:

wherein Ar¹ is an optionally substituted with one or more occurrences of—VR^(v); wherein V is a bond or is a C₁-C₆ alkylidene chain wherein upto two non-adjacent methylene units of V are independently optionallyreplaced by CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO, NRCO₂,NRCONR, SO, SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR; and each occurrenceof R^(v) is independently selected from R′, halogen, NO₂, or CN, or-QR^(x) is ═O, ═S, or ═NR′.
 31. The compound of claim 30, wherein m is 1and Ar¹ is a phenyl, pyridyl, pyrimidinyl, thiophenyl, or furanyl groupoptionally substituted with one or more occurrences of —VR^(v); whereinV is a bond or is a C₁-C₆ alkylidene chain wherein up to twonon-adjacent methylene units of V are independently optionally replacedby CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO, NRCO₂, NRCONR, SO,SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR; and each occurrence of R^(v) isindependently selected from R′, halogen, NO₂, or CN, or -QR^(x) is ═O,═S, or ═NR′.
 32. The compound of claim 27, wherein one occurrence ofWR^(y) is —CH₂N(R′)₂, and compounds have the formulae III-H-ii below,wherein m is 1-3:


33. The compound of claim 27, wherein one occurrence of WR^(y) is—N(R′)₂, and compounds have the formulae III-H-iii below, wherein m is1-3:


34. The compound of claim 27, wherein one occurrence of WR^(y) is—CON(R′)₂ and compounds have the formulae III-H-iv below, wherein m is1-3:


35. The compound of claim 32, 33, or 34, wherein —CH₂N(R′)₂ is—CH₂N(CH₂)₃, —CH₂N(CH₂CH₃)₂, —CH₂(optionally substituted N-piperazinyl),—CH₂(optionally substituted N-piperidinyl), or —CH₂(optionallysubstituted N-morpholinyl); —N(R′)₂ is —N(CH₂)₃, —N(CH₂CH₃)₂,-optionally substituted N-piperazinyl, optionally substitutedN-piperidinyl, or -optionally substituted N-morpholinyl; and —CON(R′)₂is —CON(CH₂)₃, —CON(CH₂CH₃)₂, —CO(optionally substituted N-piperazinyl),—CO(optionally substituted N-piperidinyl), or —CO(optionally substitutedN-morpholinyl).
 36. The compound of claim 32, 33, or 34, wherein—CH₂N(R′)₂ is —CH₂N(CH₂)₃, —CH₂N(CH₂CH₃)₂, —CH₂(optionally substitutedN-piperazinyl), —CH₂(optionally substituted N-piperidinyl), or—CH₂(optionally substituted N-morpholinyl); —N(R′)₂ is —N(CH₂)₃,—N(CH₂CH₃)₂, -optionally substituted N-piperazinyl, optionallysubstituted N-piperidinyl, or -optionally substituted N-morpholinyl; and—CON(R′)₂ is —CON(CH₂)₃, —CON(CH₂CH₃)₂, —CO(optionally substitutedN-piperazinyl), —CO(optionally substituted N-piperidinyl),—CO(optionally substituted N-morpholinyl), —CO(optionally substitutedpyrrolidinyl), —CO(N(H)optionally substituted pyrrolidinyl), optionallysubstituted pyrrolidinyl, or —N(H)(optionally substituted pyrrolidinyl).37. The compound of claim 21, wherein n is 0, or n is 1 and T is CH₂,—CH₂CH₂—, —CO— or —SO₂.
 38. The compound of claim 21, wherein wherein nis 0, or n is 1 and —C(H)(CH₃)—, —C(H)(CH₃)CH₂—, —CH₂C(H)(CH₃)—, or—CH₂CH₂CH₂—.
 39. The compound of claim 21, wherein n is 0, or n is 1 andT is CH₂, —CH₂CH₂—, —CO— or —SO₂—; x is 0-3; and each occurrence ofQR^(X) is independently halogen, CN, NO₂, or an optionally substitutedgroup selected from C₁₋₄alkyl, aryl, aralkyl, —N(R′)₂, —CH₂N(R′)₂, —OR′,—CH₂OR′, —SR′, —CH₂SR′, —COOR′, —NRCOR′, —CON(R′)₂, —SO₂R′, NR′SO₂R′, or—SO₂N(R′)₂.
 40. The compound of claim 1, wherein the compound isselected from:


41. The compound of claim 1, wherein the compound is selected from:


42. A composition comprising a compound of formula I:

or a pharmaceutically acceptable salt thereof, wherein: X is —(CH₂)₂—,wherein one or more of the hydrogen substituents are optionally andindependently replaced with m occurrences of —WR^(y), wherein m is aninteger selected from 0 to 4; R¹ and R², taken together with the carbonatoms to which they are bound form an optionally substituted 5-memberedheteroaryl thiophene ring, wherein the ring formed by R¹ and R² takentogether is optionally and independently substituted at one or morecarbon atoms with up to 2 occurrences of —WR^(y), each occurrence of Wis independently a bond or is a C₁-C₆ alkylidene chain wherein up to twonon-adjacent methylene units of W are independently optionally replacedby CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO, NRCO₂, NRCONR, SO,SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR; and each occurrence of R^(y) isindependently selected from R′, halogen, NO₂, or CN, or —WR^(y) is ═O,═S, or ═NR′; T is CHR′, CH₂CH(R′), —S(═O)₂, or C(═O); n is 0 or 1; Cy¹is a 3-7 membered saturated, partially unsaturated, or fully unsaturatedmonocyclic ring having 0-3 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, or an 8-10 membered saturated, partiallyunsaturated, or fully unsaturated bicyclic ring system having 0-5heteroatoms independently selected from nitrogen, oxygen, or sulfur,wherein Cy¹ is optionally and independently substituted at one or morecarbon atoms with k occurrences of -QR^(x), wherein k in an integerselected from 0 to 5; and at one or more substitutable nitrogen atomswith —R⁴; wherein Q is a bond or is a C₁-C₆ alkylidene chain wherein upto two non-adjacent methylene units of Q are independently optionallyreplaced by CO, CO₂, COCO, CONR, OCONR, NRNR, NRNRCO, NRCO, NRCO₂,NRCONR, SO, SO₂, NRSO₂, SO₂NR, NRSO₂NR, O, S, or NR; and each occurrenceof R^(X) is independently selected from R′, halogen, NO₂, or CN, or-QR^(x) is ═O, ═S, or ═NR′; each occurrence of R³ and R⁴ isindependently R′, —COR′, —CO₂(C₁₋₆aliphatic), —CON(R′)₂, or —SO₂R′; eachoccurrence of R is independently selected from hydrogen or an optionallysubstituted C₁₋₆ aliphatic group; and each occurrence of R′ isindependently selected from hydrogen or an optionally substituted groupselected from C₁₋₈ aliphatic, C₆₋₁₀ aryl, a heteroaryl ring having 5-10ring atoms, or a heterocyclyl ring having 3-10 ring atoms, or wherein Rand R′ taken together with the atom(s) to which they are bound, or twooccurrences of R′ taken together with the atom(s) to which they arcbound, form an optionally substituted 3-8 membered cycloalkyl,heterocyclyl, aryl, or heteroaryl ring having 0-3 heteroatomsindependently selected from nitrogen, oxygen, or sulfur; and the dashedbond represents a single or double bond, as valency permits; providedthat: when n is 0, and Cy¹ is 3, 4, 5-trimethoxyphenyl, and X is—(CH₂)₂—, then R¹ and R², taken together with the carbon atoms to whichthey are bound, Do not form an unsustituted thieno ring. and apharmaceutically acceptable carrier, adjuvant, or vehicle.
 43. Thecomposition of claim 42, wherein the compound is present in atherapeutically effective amount.
 44. The composition of claim 42,wherein the compound is present in an amount to detectably inhibitGSK-3, SYK, Aurora-2, CDK-2, JAK-3, LCK, SRC, and/or Tec family proteinkinase activity.
 45. The composition of claim 42, further comprising anadditional therapeutic agent selected from a chemotherapeutic oranti-proliferative agent, an anti-inflammatory agent, animmunomodulatory or immunosuppressive agent, a neurotrophic factor, anagent for treating cardiovascular disease, an agent for treatingdestructive bone disorders, an agent for treating liver disease, ananti-viral agent, an agent for treating blood disorders, an agent fortreating diabetes, or an agent for treating immunodeficiency disorders.